Epothilone derivatives and their synthesis and use

ABSTRACT

The invention relates to epothilone analog represented by formula (I) wherein (i) R 2  is absent or oxygen; “a” can be either a single or double bond; “b” can be either absent or a single bond; and “c” can be either absent or a single bond, with the proviso that if R 2  is oxygen then “b” and “c” are both a single bond and “a” is a single bond; if R 2  is absent then “b” and “c” are absent and “a” is a double bond; and if “a” is a double bond, then R 2 , “b” and “c” are absent; R 3  is a radical selected from the group consisting of hydrogen; lower alkyl; —CH═CH 2 ; —C≡CH; —CH 2 F; —CH 2 Cl; —CH 2 —OH; —CH 2 —O—(C 1 -C 6 -alkyl); and —CH 2 —S—(C 1 -C 6 -alkyl); R 4  and R 5  are independently selected from hydrogen, methyl or a protecting group; and R 1  is as defined in the specification, or a salt of a compound of formula (I) where a salt-forming group is present. A further aspect of the invention is related to the synthesis of epothilone E. These compounds have inter alia microtubuli depolymerization inhibiting activity and are e.g. useful against proliferative diseases

GOVERNMENT RIGHTS

This invention was made with government support under Grant Nos. CA46446 and CA 78045 by the National Institutes of Health. The governmenthas certain rights in the invention.

CROSS-REFERENCE

This application is a 371 of PCT/EP 99/04287 filed Jun. 21, 1999.

SUMMARY OF THE INVENTION

The present invention relates to epothilone analogs having side chainmodifications and to methods for producing such compounds, their use inthe therapy of diseases or for the manufacture of pharmaceuticalpreparations for the treatment of diseases, as well as to novelintermediates used in the synthesis of such analogs and new methods ofsynthesis.

BACKGROUND OF THE INVENTION

The epothilones (1-5) are natural substances which exhibit cytotoxicityeven against paclitaxel-resistant tumor cells by promoting thepolymerization of α- and β-tubulin subunits and stabilizing theresulting microtubule assemblies. Epothilones displace paclitaxel (theactive principle of TAXOL™) from its microtubuli binding site and arereported to be more potent than paclitaxel with respect to thestabilization of microtubules.

What is needed are analogs of epothilone A and B that exhibit superiorpharmacological properties, especially one or more of the followingproperties: an enhanced therapeutic index (e.g. a larger range ofcytotoxic doses against e.g. proliferative diseases without toxicity tonormal cells), better pharmakokinetic properties, better pharmacodynamicproperties, better solubility in water, better efficiency against tumortypes that are or become resistant to treatment with one or more otherchemotherapeutics, better properties to facilitate manufacture offormulations, e.g. better solubility in polar solvents, especially thosecomprising water, enhanced stability, convenient manufacture of thecompounds as such, improved inhibition of proliferation at the cellularlevel, high levels of microtubule stabilizing effects, and/or specificpharmacologic profiles.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to new compounds that surprisingly haveone or more of the above-mentioned advantages.

One major aspect of the invention relates to an epothilone analogcompound represented by the formula I

wherein

the waved bond indicates that bond “a” is present either in the cis orin the trans form;

(i) R₂ is absent or oxygen; “a” can be either a single or double bond;“b” can be either absent or a single bond; and “c” can be either absentor a single bond, with the proviso that if R₂ is oxygen then “b” and “c”are both a single bond and “a” is a single bond; if R₂ is absent then“b” and “c” are absent and “a” is a double bond; and if “a” is a doublebond, then R₂, “b” and “c” are absent;

R₃is a radical selected from the group consisting of hydrogen; loweralkyl, especially methyl, ethyl, n-propyl, iso-propyl, n-butyl,iso-butyl, tert-butyl, n-pentyl, n-hexyl; —CH═CH₂; —C≡CH; —CH₂F; —CH₂Cl;—CH₂—OH; —CH₂—O—(C₁-C₆-alkyl), especially —CH₂—O—CH₃; and—CH₂—S—(C₁-C₆-alkyl), especially —CH₂—S—CH₃;

R₄ and R₅ are independently selected from hydrogen, methyl or aprotecting group, preferably hydrogen; and

R₁ is a radical selected from the following structures:

 aspect of the invention, of the formula

wherein R and R′ are lower alkyl, especially methyl, or, in a broaderaspect of the invention, furthermore R′ is hydroxymethyl or fluoromethyland R is hydrogen or methyl;

(ii) and, if R₃ is lower alkyl, especially methyl, ethyl, n-propyl,iso-propyl, n-butyl, iso-butyl, tert-butyl, n-pentyl, n-hexyl; —CH═CH₂;—C≡CH; —CH₂F; —CH₂Cl; —CH₂—OH; —CH₂—O—(C₁-C₆-alkyl), especially—CH₂—O—CH₃; or —CH₂—S—(C₁-C₆-alkyl), especially —CH₂—S—CH₃, and theother symbols except R₁ have the meanings given above, R₁ can also be aradical selected from the following structures:

 or, if R₃ has one of the meanings given in the definition of R₃ aboveunder (ii) other than methyl, R₁ can also be a radical of the formula

(iii) and, if R₃ is hydrogen, lower alkyl, especially methyl, ethyl,n-propyl, iso-propyl, n-butyl, iso-butyl, tert-butyl, n-pentyl, n-hexyl;—CH═CH₂; —C≡CH; —CH₂F; —CH₂Cl; —CH₂—OH; —CH₂—O—(C₁-C₆-alkyl), especially—CH₂—O—CH₃; or —CH₂—S—(C₁-C₆-alkyl), especially —CH₂—S—CH₃, and

R₂ is oxygen, “b” and “c” are each a single bond and “a” is a singlebond, then R₁ can also be a radical of the partial formula:

(iv) and, if R₃ is lower alkyl other than methyl, especially ethyl,n-propyl, iso-propyl, n-butyl, iso-butyl, tert-butyl, n-pentyl, n-hexyl;or preferably is —CH═CH₂; —C≡CH; —CH₂F; —CH₂Cl; —CH₂—OH;—CH₂—O—(C₁-C₆-alkyl), especially —CH₂—O—CH₃; or —CH₂—S—(C₁-C₆-alkyl),especially —CH₂—S—CH₃; and the other symbols except for R₁ have themeanings given above under (i), R₁ can also be a moiety of the formula

or a salt of a compound of the formula I where a salt-forming group ispresent.

A further aspect of the invention relates to a method of synthesis of acompound of the formula

(wherein Q is hydrogen or preferably methyl) and/or a method ofsynthesis of a compound of the formula

The general terms used hereinbefore and hereinafter preferably havewithin the context of this disclosure the following meanings, unlessotherwise indicated:

The term “lower” means that the respective radical preferably has up toand including 7, more preferably up to and including 4 carbon atoms.

Lower alkyl can be linear or branched one or more times and haspreferably up to and including 7, more preferably up to and including 4carbon atoms. Preferably, lower alkyl is methyl, ethyl, n-propyl,iso-propyl, n-butyl, iso-butyl, tert-butyl or further n-pentyl orn-hexyl.

A protecting group is preferably a standard protecting group. If one ormore other functional groups, for example carboxy, hydroxy, amino, ormercapto, are or need to be protected in a compound of formulae I,because they should not take part in the reaction, these are such groupsas are usually used in the synthesis of peptide compounds, and also ofcephalo-sporins and penicillins, as well as nucleic acid derivatives andsugars.

The protecting groups may already be present in precursors and shouldprotect the functional groups concerned against unwanted secondaryreactions, such as acylations, etherifications, esterifications,oxidations, solvolysis, and similar reactions. It is a characteristic ofprotecting groups that they lend themselves readily, i.e. withoutundesired secondary reactions, to removal, typically by solvolysis,reduction, photolysis or also by enzyme activity, for example underconditions analogous to physiological conditions, and that they are notpresent in the end-products. The specialist knows, or can easilyestablish, which protecting groups are suitable with the reactionsmentioned hereinabove and hereinafter.

The protection of such functional groups by such protecting groups, theprotecting groups themselves, and their removal reactions are describedfor example in standard reference works, such as J. F. W. McOmie,“Protective Groups in Organic Chemistry”, Plenum Press, London and NewYork 1973, in T. W. Greene, “Protective Groups in Organic Synthesis”,Wiley, New York 1981, in “The Peptides”; Volume 3 (editors: E. Gross andJ. Meienhofer), Academic Press, London and New York 1981, in “Methodender organischen Chemie” (Methods of organic chemistry), Houben Weyl, 4thedition, Volume 15/I, Georg Thieme Verlag, Stuttgart 1974, in H.-D.Jakubke and H. Jescheit, “Aminosäuren, Peptide, Proteine” (Amino acids,peptides, proteins), Verlag Chemie, Weinheim, Deerfield Beach, and Basel1982, and in Jochen Lehmann, “Chemie der Kohlenhydrate: Monosaccharideund Derivate” (Chemistry of carbohydrates: monosaccharides andderivatives), Georg Thieme Verlag, Stuttgart 1974. Especially preferredprotecting groups are hydroxy protecting groups, such astert-butyldimethylsilyl or trityl.

R₄ and R₅ are preferably hydrogen.

The waved bond starting from the carbon atom bearing R₃ means that bond“a” is present in the trans- or preferably the cis-form.

Salts are especially the pharmaceutically acceptable salts of compoundsof formula I.

Such salts are formed, for example, as acid addition salts, preferablywith organic or inorganic acids, from compounds of formula I with abasic nitrogen atom, especially the pharmaceutically acceptable salts.Suitable inorganic acids are, for example, halogen acids, such ashydrochloric acid, sulfuric acid, or phosphoric acid. Suitable organicacids are, for example, carboxylic, phosphonic, sulfonic or sulfamicacids, for example acetic acid, propionic acid, octanoic acid, decanoicacid, dodecanoic acid, glycolic acid, lactic acid, fumaric acid,succinic acid, adipic acid, pimelic acid, suberic acid, azelaic acid,maleic acid, tartaric acid, citric acid, amino acids, such as glutamicacid or aspartic acid, maleic acid, hydroxymaleic acid, methylmaleicacid, cyclohexanecarboxylic acid, adamantanecarboxylic acid, benzoicacid, salicylic acid, 4-aminosalicylic acid, phthalic acid, phenylaceticacid, mandelic acid, cinnamic acid, methane- or ethane-sulfonic acid,2-hydroxyethanesulfonic acid, ethane-1,2-disulfonic acid,benzenesulfonic acid, 2-naphthalenesulfonic acid,1,5-naphthalenedisulfonic acid, 2-, 3- or 4-methylbenzenesulfonic acid,methylsulfuric acid, ethylsulfuric acid, dodecylsulfuric acid,N-cyclohexylsulfamic acid, N-methyl-, N-ethyl- or N-propyl-sulfamicacid, or other organic protonic acids, such as ascorbic acid.

For isolation or purification purposes it is also possible to usepharmaceutically unacceptable salts, for example picrates orperchlorates. For therapeutic use, only pharmaceutically acceptablesalts or free compounds are employed (where applicable in the form ofpharmaceutical preparations), and these are therefore preferred.

In view of the close relationship between the novel compounds in freeform and those in the form of their salts, including those salts thatcan be used as intermediates, for example in the purification oridentification of the novel compounds, any reference to the freecompounds hereinbefore and hereinafter is to be understood as referringalso to the corresponding salts, as appropriate and expedient.

The term “about” in connection with numerical values, e.g. “about 2-foldmolar excess” or the like, is preferably intended to mean that the givennumerical value may deviate from the given number by up to ±10%, morepreferably by up to ±3%; most preferably, the numerical value is exactlyas given.

In a preferred embodiment of the invention, the compounds of formula Ias described under (iv) above

are excluded from the scope of the invention.

Also, a group of compounds of the formula I without a compound of theformulae I wherein R₁ is a moiety of any one of the formulae

is preferred (the remaining symbols having the meanings defined for acompound of the formula I).

Especially preferred is either a free compound of the formula I, or asalt thereof.

Bioactivity: The compound(s) of the invention can be used for thetreatment of a proliferative disease, especially a cancer, like cancersof the lung, especially non-small lung cell lung carcinoma, of theprostate, of the intestine, e.g. colorectal cancers, epidermoid tumors,such as head and/or neck tumors, or breast cancer, or other cancers likecancers of the bladder, pancreas or brain or melanoma, includingespecially the treatment of cancers that are multidrug-resistant (e.g.due to the expression of p-glycoprotein=P-gp) and/or refractory totreatment with paclitaxel (e.g. in the form of TAXOL).

Biological Evaluation

The ability of the compounds of the present invention to block thedepolymerization of microtubuli can be shown by the following assay:

Microtubule assays are carried out following literature procedures andevaluate synthesized compounds for their ability to form and stabilizemicrotubules. Cytotoxicity studies are carried out as well.

The compounds of formula I are tested for their action on tubulinassembly using purified tubulin with an assay developed to amplifydifferences between compounds more active than Taxol. Compounds of theformula I are found to have a high level of cytotoxic and tubulinpolymerization activity, as compared to Epothilones A and B. (Lin et al.Cancer Chemother. Pharmacol. 38, 136-140 (1996); Rogan et al. Science244, 994-996 (1984)).

Filtration Colorimetric Assay

Microtubule protein (0.25 ml of 1 mg/ml) is placed into an assay tubeand 2.5 μl of the test compound are added. The sample is mixed andincubated at 37° C. for 30 min. Sample (150 μl) is transferred to a wellin a 96-well Millipore Multiscreen Durapore hydrophilic 0.22 μm poresize filtration plate which has previously been washed with 200 μl ofMEM buffer under vacuum. The well is then washed with 200 μl of MEMbuffer. To stain the trapped protein on the plate, 50 μl amido blacksolution [0.1% naphthol blue black (Sigma)/45% methanol/10% acetic acid]are added to the filter for 2 min; then the vacuum is reapplied. Twoadditions of 200 μl amido black destain solution (90% methanol/2% aceticacid) are added to remove unbound dye. The signal is quantitated by themethod of Schaffner and Weissmann et al. Anal. Biochem., 56: 502-514,1973 as follows: 200 μl of elution solution (25 mM NaOH-0.05 mm EDTA-50%ethanol) are added to the well and the solution is mixed with a pipetteafter 5 min. Following a 10-min incubation at room temperature, 150 μlof the elution solution are transferred to the well of a 96-well plateand the absorbance is measured on a Molecular Devices Microplate Reader.

Cytotoxicity experiments with 1A9, 1A9PTX10 (α-tubulin mutant), and1A9PTX22 (α-tubulin mutant) cell lines can reveal the cytotoxic activityof the compounds of formula I. Like the naturally occurring epothilones1 and 2, compounds of the formula I show significant activity againstthe altered α-tubulin-expressing cell lines 1A9PTX10 and 1A9PTX22. Forcompounds of the formula I, the preferred IC50 values (concentrationwhere half-maximal growth inhibition of tumor cells is found incomparison with a control without added inhibitor of the formula I) canlie in the range of 1 to 1000 nM, preferably from 1 to 200 nM.

The ability of the compounds of the present invention to inhibit tumorgrowth can be shown by the following assays with the following celllines:

Colorimetric Cytotoxicity Assay for Anticancer-Drug Screening

The calorimetric cytotoxicity assay used is adapted from Skehan et al(Journal of National Cancer Inst 82:1107-1112, 19901). The procedureprovides a rapid, sensitive, and inexpensive method for measuring thecellular protein content of adherent and suspension cultures in 96-wellmicrotiter plates. The method is suitable for the National CancerInstitute's disease-oriented in vitro anticancer-drug discovery screen.

In particular, cultures fixed with trichloroacetic acid are stained for30 minutes with 0.4% (wt/vol) sulforhodamine B (SRB) dissolved in 1%acetic acid. Unbound dye is removed by four washes with 1% acetic acid,and protein-bound dye is extracted with 10 mM unbuffered Tris base[tris(hydroxymethyl)aminomethane] for determination of optical densityin a computer-interfaced, 96-well microtiter plate reader. The SRB assayresults are linear with the number of cells and with values for cellularprotein measured by both the Lowry and Bradford assays at densitiesranging from sparse subconfluence to multilayered supraconfluence. Thesignal-to-noise ratio at 564 nm is approximately 1.5 with 1,000 cellsper well.

The SRB assay provides a calorimetric end point that is nondestructive,indefinitely stable, and visible to the naked eye. It provides asensitive measure of drug-induced cytotoxicity. SRB fluoresces stronglywith laser excitation at 488 nm and can be measured quantitatively atthe single-cell level by static fluorescence cytometry (Skehan et al(Journal of National Cancer Inst 82:1107-1112, 19901)).

Alternatively, the efficiency of the compounds of the formula I asinhibitors of microtubuli depolymerisation can be demonstrated asfollows:

Stock solutions of the test compounds are made in DMSO and stored at−20° C. Microtubuli-protein is obtained from pig brain by two cycles oftemperature dependent depolymerisation/polymerisation, as described (seeWeingarten et al., Biochemistry 1974; 13: 5529-37). Working stocksolutions of microtubule protein (meaning tubulin plusmicrotubuli-associated proteins) are stored at −70° C. The degree of themicrotubuli protein polymerisation induced by a test compound ismeasured essentially as known from the literature (see Lin et al.,Cancer Chem. Pharm. 1996; 38:136-140). In short, 5 μl stock solution ofthe test compound are pre-mixed in the twenty-fold of the desired finalconcentration with 45 μl of water at room temperature, and the mixtureis then placed on ice. An aliquot of the working stock solution of pigbrain microtubuli protein is thawed quickly and then diluted to 2 mg/mlwith ice-cold 2× MEM buffer (200 ml MES, 2 mM EGTA, 2 mM MgCl₂, pH 6.7)(MES=2-morpholinoethanesulfonic acid,EGTA=ethylenglycol-bis-(2(2-aminoethyl)-tetraacetic acid). Thepolymerisation reaction is then started by the addition of each time 50μl diluted microtubuli-protein to the test compound, followed byincubation of the sample in a water bath with room temperature. Then thereaction mixtures are placed in an Eppendorf microcentrifuge andincubated for additional 15 min at room temperature. The samples arethen centrifuged for 20 min at 14000 rpm at room temperature forseparating polymerized from non-polymerized microtubuli protein. Asindirect measure for the tubulin-polymerisation the proteinconcentration of the supernatant (which contains the rest of theun-polymerised, soluble microtubuti protein) is determined according tothe Lowry method (DC Assay Kit, Bio-Rad Laboratories, Hercules, Calif.),and the optical density (OD) of the color reaction is determined at 750nm with a spectrometer (SpectraMax 340, Molecular Devices, Sunnyvale,Calif.). The differences in the OD's between samples treated with a testcompound and vehicle-treated controls are compared with those of testincubations which contain 25 μM Epothilon B (positive controls). Thedegree of polymerisation that is induced by a test compound is expressedrelatively to the positive controls (100%). By comparison of severalconcentrations the EC50 (concentration where 50% of the maximalpolymerisation is found) can be determined. For compounds of the formulaI the EC50 lies preferably in the range of 1 to 200, preferably 1 to 50μM. The induction of tubulin polymerisation of test compound of theformula I in 5 μM concentration as parentage in comparison to 25 μMepothilone B preferably lies in the range of 50 to 100%, especially 80to 100%.

The efficiency against tumor cells can also be shown in the followingway:

Stock solutions of the test compound of formula I 10 mM) in DMSO areprepared and stored at −20° C. Human KB-31 and (multidrug-resistant,P-gp 170 overexpressing) KB-8511 epidermoid carcinoma cells are from Dr.M. Baker, Roswell Park Memorial Institute (Buffalo, N.Y., USA) (fordescription see also Akiyama et al., Somat. Cell. Mol. Genetics 11,117-126 (1985) und Fojo A., et al., Cancer Res. 45, 3002-3007(1985)—KB-31 and KB-8511 both are derivatives of the KB-cell line(American Type Culture Collection) and are human epidermoid carcinomacells. KB-31 cells can be cultivated in mono-layers using calf serum(M.A. Bioproducts), L-glutamine (Flow), penicillin (50 Units/ml) andstreptomycin (50 μg/ml (Flow); they then grow with a doubling rate ofabout 22 hours, and the relative efficiency of plating them out lies atabout 60%. KB-8511 is a variant derived from the KB-31 cell line whichhas been obtained by treatment cycles with colchicine, and it shows anabout 40-fold relative resistance against colchicin in comparison toKB-31 cells). The cells are incubated at 37° C. in an incubator with 5%v/v CO₂ and at 80% relative atmospheric humidity in MEM Alpha-mediumwhich contains ribonucleosides and desoxyribonucleosides (Gibco BRL),complemented with 10 IU Penicillin, 10 μg/ml Streptomycin and 5% fetalcalf serum. The cells are spread in an amount of 1.5×10³ cells/well in96-well-microtiter plates and incubated overnight. Serial dilutions ofthe test compounds in culture medium are added at day 1. The plates arethen incubated for an additional period of four days, after which thecells are fixed using 3.3% v/v glutaraldehyde washed with water andfinally stained with 0.05% w/v methylen blue. After washing again, thestain is eluted with 3% HCl and the optical density at 665 nm ismeasured with a SpectraMax 340 (Molecular Devices, Sunnyvale, Calif.).IC50-values are determined by mathematically fitting the data to curvesusing the SoftPro2.0 program (Molecular Devices, Sunnyvale, Calif.) andthe formula

[(OD treated)−(OD start)]/[(OD control)−(OD start)]×100.

The IC50 is defined as the concentration of a test compound at the endof the incubation period that leads to 50% of the number of cells incomparison to controls without test compound (concentration athalfmaximal inhibition of cell growth). Compounds of the formula Ipreferably show here and IC50 in the range from 0.1×10⁻⁹ to 500×10⁻⁹ M,preferably between 0.1 and 60 nM.

Comparable testing can also be made with other tumor cell lines, such asA459 (lung; ATCC CCL 185), NCIH460 (lung), Colo 205 (colon; ATCC No. CCL222) (HCT-15 (colon; ATCC CCL 225—ATCC=American Type Culture Collection(Rockville, Md., USA)), HCT-116 (colon), Du145 (prostate; ATCC No. HTB81; see also Cancer Res. 37, 4049-58 [1978]), PC-3M(prostate-hormone-insensitive derivative obtained from Dr. I. J. Fidler(M D Anderson Cancer Center, Houston, Tex., USA) and derived from PC-3that is a cell line available from ATCC (ATCC CRL 1435)), MCF-7 (breast;ATCC HTB 22) or MCF-7/ADR (breast, multidrug resistant; for descriptionsee Blobe G. C.et al., J. Biol. Chem. (1983), 658-664; the cell line ishighly resistant (360- to 2400-fold) to doxorubicin and Vinca alkaloidscompared over MDR-7 “wild type” cells)), where similar results areobtained as with KB-31 and KB-8511 cells. Compounds of the formula Ipreferably show here and IC50 in the range from 0.1×10⁻⁹ to 500×10⁻⁹ M,preferably between 0.1 and 60 nM.

Based on these properties, the compounds of the formula I (meaining alsosalts thereof) are appropriate for the treatment of proliferativediseases, such as especially tumor diseases, including also metastasiswhere present, for example of solid tumors, such as lung tumor, breasttumor, colorectal cancer, prostate cancer, melanoma, brain tumor,pancreas tumor, head-and-neck tumor, bladder cancer, neuroblastoma,pharyngeal tumor, or also of proliferative diseases of blood cells, suchas leukaemia; or further for the treatment of other diseases thatrespond to treatment with microtubuli depolymerisation inhibitors, suchas psoriasis. The compounds of formula I, or salts thereof, are alsoappropriate for covering medical implants (useful in prophylaxis ofrestenosis) (see WO 99/16416, priority Sep. 29, 1997).

The in vivo activity of a compound of the invention can be demonstratedwith the following animal model:

Female or male BALB/c nu/nu (nude) mice are kept under sterileconditions (10 to 12 mice per Type III cage) with free access to foodand water. Mice weigh between 20 and 25 grams at the time of tumorimplantation. Tumors are established by subcutaneous injection of cells(minimum 2×10⁶ cells in 100 μl PBS or medium) in carrier mice (4-8 miceper cell line). The resulting tumors are serially passaged for a minimumof three consecutive transplantations prior to start of treatment. Tumorfragments (approx. 25 mg) are implanted s.c. into the left flank ofanimals with a 13-gauge trocar needle while the mice are exposed toForene (Abbott, Switzerland) anesthesia.

Tumor growth and body weights are monitored once or twice weekly. Alltreatments are administered intravenously (i.v.) and are initiated whena mean tumor volume of approximately 100 to 250 mm³ is attained,depending upon the tumor type. Tumor volumes are determined using theformula (L×D×π)/6 (see Cancer Chemother. Pharmacol. 24:148-154, [1989]).Treatments with epothilones of the formula I vary the dose and thefrequency of administration. Comparator agents are administeredaccording to previously determined optimal treatment regimens. Inaddition to presenting changes in tu-mor volumes over the course oftreatment, antitumor activity is expressed as T/C% (mean increase oftumor volumes of treated animals divided by the mean increase of tumorvolu-mes of control animals multiplied by 100). Tumor regression (%)represents the smallest mean tumor volume compared to the mean tumorvolume at the start of treatment, accor-ding to the formula Regression(%)=(1−Vend/Vstart)×100 (Vend=final mean tumor volume, Vstart=mean tumorvolume at the start of treatment.

With this model, the inhibitory effect of a compound of the invention ongrowth e.g. of tumors derived from the following cell lines can betested:

Human colorectal adenocarcinoma cell line HCT-15 (ATCC CCL 225) is fromthe American Type Culture Collection (Rockville, Md., USA), and thecells are cultivated in vitro as recommended by the supplier. HCT-15 isan epithelial-like cell line (Cancer Res. 39: 1020-25 [1979]) that ismulti-drug resistant by virtue of over-expression of P-glycoprotein(P-gp, gp170, MDR-1; Anticancer Res. 11: 1309-12 [1991]; J. Biol. Chem.264: 18031-40 [1989]; Int. J. Cancer 1991; 49: 696-703 [1991]) andglutathione-dependent resistance mechanisms (Int. J. Cancer 1991; 49:688-95.[1991]). The Colo 205 cell line is also a human colon carcinomacell line (ATCC No. CCL 222; see also Cancer Res. 38, 1345-55 [1978]which was isolated from ascitic fluid of a patient, dis-playsepithelial-like morphology and is generally considered to bedrug-sensitive. A human androgen-independent prostate cancer cell lineis used to establish subcutaneous and orthotopic models in mice. Thehuman metastatic prostate carcinoma PC-3M is obtained from Dr. I. J.Fidler (M D Anderson Cancer Center, Houston, Tex., USA) and is culturedin Ham's F12K media supplemented with 7% v/v FBS. The PC-3M cell line isthe result of isolation from liver metastasis produced in nude micesubsequent to intrasplenic injection of PC-3 cells [ATCC CRL 1435;American Type Culture Collection (Rockville, Md., USA)], and they cangrow in Eagle's MEM supplemented with 10% fetal bovine serum, sodiumpyruvate, non-essential amino acids, L-glutamine, a two-fold vitaminsolution (Gibco Laboratories, Long Island, N.Y.) andpenicillin-streptomycin (Flow Laboratories, Rockville, Md.). The PC-3Mcell line is hormone-insensitive (that is, it grows in the absence ofandrogens). The PC-3 cell line is androgen receptor negative, as ispresumably the derived PC-3M cell line. PC-3 is a cell line availablefrom ATCC (ATCC CRL 1435) and corresponds to a grade IV prostaticadenocarcinoma isolated from a 62-year-old Caucasian male; the cellsexhibit low acid phosphatase and testosterone-5-a-reductase activity.The cells are near-triploid with a modal number of 62 chromosomes. Nonormal Y chromosomes can be detected by Q-band analysis. Human lungadenocarcinoma A549 (ATCC CCL 185; isolated as explant culture from lungcarcinoma tissue from a 58-year-old Caucasian male); shows epithelialmorphology and can synthesize lecithin with a high percentage ofdesaturated fatty acids utilizing the cytidine diphosphocholine pathway;a subtelocentric marker chromosome involving chromosome 6 and the longarm of chromosome 1 is found in all metaphases. The human breastcarcinoma ZR-75-1 (ATCC CRL 1500; isolated from a malignant asciticeffusion of a 63-year-old Caucasian female with infiltrating ductalcarcinoma); is of mammary epithelial origin; the cells possess receptorsfor estrogen and other steroid hormones and have a hypertriploidchromosome number. The human epidermal (mouth) carcinoma cell lineKB-8511 (a P-gp over-expressing cell line derived from the epidermoid(mouth) KB-31 carcinoma cell line) is obtained from Dr. R. M. Baker,Roswell Park Memorial Institute (Buffalo, N.Y., USA) (for descriptionsee Akiyama et al., Somat. Cell. Mol. Genetics 11, 117-126 (1985) andFojo A., et al., Cancer Res. 45, 3002-3007 (1985)) and is cultured aspreviously described (Meyer, T., et al., Int. J. Cancer 43, 851-856(1989)). KB-8511 cells, like KB-31, are derived from the KB cell line(ATCC) and they are human epidermal carcinoma cells; KB-31 cells can begrown in mono-layer using Dulbecco's modified Eagle's medium (D-MEM)with 10% fetal calf serum (M.A. Bioproducts), L-glutamine (Flow),penicillin (50 units/ml) and streptomycin (50 mg/ml (Flow); they thengrow with a doubling time of 22 h, and their relative plating efficiencyis approximately 60%. KB-8511 is a cell line derived from the KB-31 cellline by use of colchicine treatment cycles; it shows about a 40-foldrelative resistance against colchicine when compared with the KB-31cells; it can be grown under the same conditions as KB-31.”

Solubility: The water solubility is determined as follows, for example:the compounds of formula I, or the salts thereof, are stirred with waterat room temperature until no further compound dissolves (about 1 hour).The solubilities found are preferably between 0.01 and 1% by weight.

Within the groups of preferred compounds of formula I mentionedhereinafter, definitions of substituents from the general definitionsmentioned hereinbefore may reasonably be used, for example, to replacemore general definitions with more specific definitions or especiallywith definitions characterised as being preferred.

The invention preferably relates to a compound of the formula I wherein

R₂ is absent or oxygen; “a” can be either a single or double bond; “b”can be either absent or a single bond; and “c” can be either absent or asingle bond, with the proviso that if R₂ is oxygen then “b” and “c” areboth a single bond and “a” is a single bond; if R₂ is absent then “b”and “c” are absent and “a” is a double bond; and if “a” is a doublebond, then R₂, “b” and “c” are absent;

R₃ is a radical selected from the group consisting of hydrogen; loweralkyl, especially methyl, ethyl, n-propyl, iso-propyl, n-butyl,iso-butyl, tert-butyl, n-pentyl, n-hexyl; —CH═CH₂; —C≡CH; —CH₂F; —CH₂Cl;—CH₂—OH; —CH₂—O—(C₁-C₆-alkyl), especially —CH₂—O—CH₃; and—CH₂—S—(C₁-C₆-alkyl), especially —CH₂—S—CH₃;

R₄ and R₅ are independently selected from hydrogen, methyl or aprotecting group, preferably hydrogen; and

R₁ is a radical selected from the following structures:

wherein R and R′ are lower alkyl, especially methyl;

or a salt thereof where salt-forming groups are present.

The invention preferably also relates to a compound of the formula Iwherein

R₂ is absent or oxygen; “a” can be either a single or double bond; “b”can be either absent or a single bond; and “c” can be either absent or asingle bond, with the proviso that if R₂ is oxygen then “b” and “c” areboth a single bond and “a” is a single bond; if R₂ is absent then “b”and “c” are absent and “a” is a double bond; and if “a” is a doublebond, then R₂, “b” and “c” are absent;

R₃ is a radical selected from the group consisting of hydrogen; loweralkyl, especially methyl, ethyl, n-propyl, iso-propyl, n-butyl,iso-butyl, tert-butyl, n-pentyl, n-hexyl; —CH═CH₂; —C≡CH; —CH₂F; —CH₂Cl;—CH₂—OH; —CH₂—O—(C₁-C₆-alkyl), especially —CH₂—O—CH₃; and—CH₂—S—(C₁-C₆-alkyl), especially —CH₂—S—CH₃;

R₄ and R₅ are independently selected from hydrogen, methyl or aprotecting group, preferably hydrogen; and

R₁ is a radical selected from the following structures:

wherein R′ is hydroxymethyl or fluoromethyl and R is hydrogen or methyl;

or a salt thereof where a salt-forming group is present.

The invention preferably also relates to a compound of the formula Iwherein

R₂ is absent or oxygen; “a” can be either a single or double bond; “b”can be either absent or a single bond; and “c” can be either absent or asingle bond, with the proviso that if R₂ is oxygen then “b” and “c” areboth a single bond and “a” is a single bond; if R₂ is absent then “b”and “c” are absent and “a” is a double bond; and if “a” is a doublebond, then R₂, “b” and “c” are absent;

R₃ is a radical selected from the group consisting of lower alkyl,especially methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl,tert-butyl, n-pentyl, n-hexyl; —CH═CH₂; —C≡CH; —CH₂F; —CH₂Cl; —CH₂—OH;—CH₂—O—(C₁-C₆-alkyl), especially —CH₂—O—CH₃; and —CH₂—S—(C₁-C₆-alkyl),especially —CH₂—S—CH₃,

R₄ and R₅ are independently selected from hydrogen, methyl or aprotecting group, preferably hydrogen; and

R₁ is a radical selected from the following structures:

or a salt thereof where one or more salt-forming groups are present.

The invention preferably also relates to a compound of the formula Iwherein

R₂ is absent or oxygen; “a” can be either a single or double bond; “b”can be either absent or a single bond; and “c” can be either absent or asingle bond, with the proviso that if R₂ is oxygen then “b” and “c” areboth a single bond and “a” is a single bond; if R₂ is absent then “b”and “c” are absent and “a” is a double bond; and if “a” is a doublebond, then R₂, “b” and “c” are absent;

R₃ is a radical selected from the group consisting of lower alkyl otherthan methyl, especially ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl,tert-butyl, n-pentyl, n-hexyl; —CH═CH₂; —C≡CH; —CH₂F; —CH₂Cl; —CH₂—OH;—CH₂—O—(C₁-C₆-alkyl), especially —CH₂—O—CH₃; and —CH₂—S—(C₁-C₆-alkyl),especially —CH₂—S—CH₃,

R₄ and R₅ are independently selected from hydrogen, methyl or aprotecting group, preferably hydrogen; and

R₁ is a radical of the formula

or a salt thereof if one or more salt-forming groups are present.

The invention preferably also relates to a compound of the formula Iwherein

R₂ is oxygen, “b” and “c” are each a single bond and “a” is a singlebond,

R₃ is a radical selected from the group consisting of lower alkyl,especially methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl,tert-butyl, n-pentyl, n-hexyl; —CH═CH₂; —C≡CH; —CH₂F; —CH₂Cl; —CH₂—OH;—CH₂—O—(C₁-C₆-alkyl), especially —CH₂—O—CH₃; and —CH₂—S—(C₁-C₆-alkyl),especially —CH₂—S—CH₃,

R₄ and R₅ are independently selected from hydrogen, methyl or aprotecting group, preferably hydrogen; and

R₁ is a radical selected from the group consisting of the followingstructures:

or a salt thereof where one or more salt-forming groups are present.

The invention preferably also relates to a compound of the formula Iwherein

R₂ is absent or oxygen; “a” can be either a single or double bond; “b”can be either absent or a single bond; and “c” can be either absent or asingle bond, with the proviso that if R₂ is oxygen then “b” and “c” areboth a single bond and “a” is a single bond; if R₂ is absent then “b”and “c” are absent and “a” is a double bond; and if “a” is a doublebond, then R₂, “b” and “c” are absent;

R₃ is a radical selected from the group consisting of lower alkyl otherthan methyl, especially ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl,tert-butyl, n-pentyl or n-hexyl; —CH═CH₂; —C≡CH; —CH₂F; —CH₂Cl; —CH₂—OH;—CH₂—(C₁-C₆-alkyl), especially —CH₂—O—CH₃; and —CH₂—S—(C₁-C₆-alkyl),especially —CH₂—S—CH₃,

R₄ and R₅ are independently selected from hydrogen, methyl or aprotecting group, preferably hydrogen; and

R₁ is a radical of the formula

or a salt of a compound of the formula I where a salt-forming group ispresent.

More preferably, the invention relates to a compound of the formula Ia,

wherein, independent of each other, the R moieties are hydrogen ormethyl, or a salt thereof.

More preferably, the invention also relates to a compound of the formulaIb,

wherein, independent of each other, the R moieties are hydrogen ormethyl, or a salt thereof.

The invention most specifically also relates to a compound of theformula Ic

wherein R* is methyl.

The invention most specifically also relates to a compound of theformula Id

wherein A is ethyl, fluoromethyl, methoxy, methylthio or ethenyl(—CH═CH₂) and D is hydrogen, fluoro, hydroxy or methyl, especiallyhydrogen.

The invention most specifically also relates to a compound of theformula Ie

wherein A is ethyl, fluoromethyl, methoxy, methylthio or ethenyl(—CH═CH₂) and D is hydrogen, fluoro, hydroxy or methyl.

The invention most specifically relates to the compounds of the formulaI given in the examples, or the pharmaceutically acceptable saltsthereof where one or more salt-forming groups are present.

Most preferably, the invention relates to a compound selected from thegroup consisting of compound D (Example 1), the compound of Example 2A), the compound of Example 2C, compound 18b (see Example 4), compound19b (see Example 4), compound 46 (see Example 4), compound 50 (seeExample 4), compound 52 (see Example 4), compound 53 (see Example 4),compound 58 (see Example 4), compound 59 (see Example 4), compound 66(see Example 4), compound 67 (see Example 4), and compound 68 (seeExample 4), or a pharmaceutically acceptable salt thereof if one or moresalt-forming groups are present.

The compounds of the invention can be synthesized using methods inanalogy to methods that are known in the art, preferably by a methodcharacterized by

a) reacting a iodide of the formula II,

wherein R₂, R₃, R₄, R₅, a, b and c and the waved bond have the meaningsgiven under formula I, with a tin compound of the formula III,

R₁—Sn(R)₃  (III)

wherein R₁ has the meanings given under formula I and R is lower alkyl,especially methyl or n-butyl, or

b) reacting a tin compound of the formula IV,

wherein R₂, R₃, R₄, R₅, a, b and c and the waved bond have the meaningsgiven under formula I, with a iodide of the formula V,

R₁—I  (V)

wherein R₁ has the meanings given under formula I;

and, if desired, a resulting compound of the formula I is converted intoa different compound of the formula I, a resulting free compound of theformula I is converted into a salt of a compound of the formula I,and/or a resulting salt of a compound of the formula I is converted intoa free compound of the formula I or into a different salt of a compoundof the formula I, and/or a stereoisomeric mixture of compounds offormula I is separated into the corresponding isomers.

DETAILED DESCRIPTION OF THE PREFERRED PROCESS CONDITIONS

In all starting materials, where required, functional groups that shallnot participate in the reaction are protected by protecting groups,especially standard protecting groups. The protecting groups, theirintroduction and their cleavage are known in the art, for example, theyare described in the standard references mentioned above.

Reaction a): The reaction (a (preferably improved) Stille coupling)preferably takes place under standard conditions; more preferably, thereaction takes place

(i) in an appropriate solvent, e.g. toluene, at elevated temperature,especially about 90 to about 100° C., preferably with an excess of thetin compound of the formula III, preferably in the 1.1- to 3-, e.g. the1.5- to 2-fold molar excess; and a catalytic amount, preferably of about1 to 30%, preferably 5 to 10%, of Pd(PPh₃)₄; or

(ii) in an appropriate solvent, e.g. dimethylformamide (DMF), attemperatures of from 10 to 40° C., especially at 25° C., preferably withan excess of the tin compound of the formula III, preferably in the 1.1-to 3-, e.g. the 1.5- to 2.3-fold molar excess; in the presence of acatalytic amount, preferably of 10 to 50%, especially 20 to 30%, ofPd(MeCN)₂Cl₂. Alternative conditions for this coupling also comprise theuse of the following reagents and/or conditions:

(iii). cuprous 2-thiophene carboxylate, N-methyl-2-pyrrolidine.

(iv). PdCl₂(MeCN)₂ (cat.), DMF, 50-150° (with or without additon oftertiary base).

(v). Pd(PPh₃)₄/Cul (cat), DMF, 50-150° (with or without addition oftertiary base)

Reaction b): The reaction (an improved Stille coupling) preferably takesplace under standard conditions; more preferably, the reaction takesplace in an appropriate solvent, especially DMF, at temperatures of from50 to 100, preferably from 80 to 85° C., preferably with an excess ofthe iodide of the formula V, in the presence of a catalytic amount ofAsPh₃, preferably about 0.4 equivalents, Cul, preferably about 0.1equivalents, and PdCl₂(MeCN)₂, preferably about 0.2 equivalents.

Especially preferred are the reaction conditions mentioned in theexamples.

Conversions of Compounds/salts

Compounds of the formula I can be converted into different compounds offormula I by standard or novel methods.

For example, a compound of the formula I wherein R₂ is absent, b and care absent and a is a double bond, and the other moieties are asdescribed for compounds of the formula I, can be converted into thecorresponding epoxide wherein R₂ is O and b and c are present while a isa single bond. Preferably, the epoxidation takes place in the presenceof (+)-diethyl-D-tartrate ((+)-DET) (preferably about 0.5 equivalents),Ti(i-PrO)₄ (preferably about 0.5 equivalents), tert-butylhydroperoxide(preferably about 2.2 equivalents) and molecular sieve, especially 4 Åmolecular sieves, in an appropriate solvent, e.g. methylene chloride andoptionally an alkane, such as decane, at low temperatures, preferably-78 to 0° C., especially about −40° C.; or in presence of hydrogenperoxide (preferably about 30 equivalents), acetonitrile (preferablyabout 60 equivalents), a base, especially KHCO₃ (preferably about 10equivalents) in an appropriate solvent, e.g. an alcohol, preferablymethanol, at preferred temperatures between 10 and 40° C., e.g. at about25° C.

A compound of the formula I wherein R₃ is hydroxymethyl can be convertedinto a compound of formula I wherein R₃ is fluoromethyl, e.g. bytreatment with DAST (preferably 1.05 to 1.4 equivalents) in anappropriate solvent, e.g. methylene chloride, at low temperatures,preferably at −95 to 0° C., especially at about −78° C. DAST isdiethylamino-sulfur trifluoride.

A compound of the formula I wherein R₃ is iodomethyl can be convertedinto a compound of formula I wherein R₃ is methyl, e.g. by treatmentwith cyanoborohydride (preferably about 10 equivalents) in HMPA(hexamethylphosphoric triamide) at elevated temperatures, e.g. at 40 to45° C.

Other conversions can be made in accordance with known procedures, e.g.those given in PCT application WO 98/25929, which is herewithincorporated by reference.

Salts of a compound of formula I with a salt-forming group may beprepared in a manner known per se. Acid addition salts of compounds offormula I may thus be obtained by treatment with an acid or with asuitable anion exchange reagent.

Salts can usually be converted to free compounds, e.g. by treating withsuitable basic agents, for example with alkali metal carbonates, alkalimetal hydrogencarbonates, or alkali metal hydroxides, typicallypotassium carbonate or sodium hydroxide.

The resulting free compounds can then, if desired, be converted intodifferent salts as described for the formation of salts from the freecompounds.

Stereoisomeric mixtures, e.g. mixtures of diastereomers, can beseparated into their corresponding isomers in a manner known per se bymeans of suitable separation methods. Diastereomeric mixtures forexample may be separated into their individual diastereomers by means offractionated crystallization, chromatography, solvent distribution, andsimilar procedures. This separation may take place either at the levelof a starting compound or in a compound of formula I itself. Enantiomersmay be separated through the formation of diastereomeric salts, forexample by salt formation with an enantiomer-pure chiral acid, or bymeans of chromatography, for example by HPLC, using chromatographicsubstrates with chiral ligands.

Starting Materials

Starting materials and intermediates are known in the art, commerciallyavailable, and/or prepared in accordance with methods known in the artor in analogy thereto.

Compounds of the formula II and of the formula III can, for example, besynthesized as described in PCT application WO 98/25929, which isherewith incorporated by reference, or as described or in analogy to themethods in the examples.

Compounds of the formula IV are accessible by reaction of the respectivecompounds of the formula II, for example by reaction of a compound ofthe formula II with (R)₆Sn₂, wherein R is lower alkyl, especially methylor n-butyl, in the presence of an appropriate nitrogen base, e.g.Hünig's base, and in the presence of catalytic amount (preferably about0.1 equivalents) of Pd(PPh₃)₄ in an appropriate solvent, e.g. toluene,at elevated temperatures, e.g. 30 to 90° C., especially 80 to 85° C.lodides of the formula V are known and can be obtained according toliterature procedures, or they are commercially available. For example,2-iodo-6-methyl pyridine can be obtained according to Klei, E.; Teuben,J. H. J. Organomet. Chem. 1981, 214, 53-64; 2-iodo-5-methyl pyridineaccording to Talik, T.; Talik, Z. Rocz. Chem. 1968, 42, 2061-76; and2-iodo-4-methyl pyridine according to Talik, T.; Talik, Z. Rocz. Chem.1968, 42, 2061-76, Yamamoto, Y.; Yanagi, A. Heterocycles 1981, 16,1161-4 or Katritzky, A. R.; Eweiss, N. F.; Nie, P.-L. JCS, Perkin TransI 1979, 433-5. The corresponding hydroxymethyl-substituted compounds offormula V are available for example by oxidation of the methyl groups ofthe iodides mentioned above with SeO₂ and subsequent reduction, e.g.with NaBH₄ or DIBALH) of the aldehyde or by oxidation of the methylgroup to form the acid (for example with KMnO₄) and subsequent reductionof the ester e.g. with DIBAL.

Preferably, new or also known starting materials and intermediates canbe prepared in accordance with or in analogy to the methods described inthe examples, where the amounts, temperatures and the like of therespective reactions can be modified, e.g. by varying in the range of±99%, preferably ±25%, and other appropriate solvents and reagents canbe used.

The invention relates also to all new intermediates, especially thosementioned in the Examples.

The invention also relates to a method of synthesis of a compound of theformula VI

which is characterized in that a compound of the formula VII

wherein R₃ is lower alkyl, especially methyl or n-butyl, is coupled witha iodide of the formula VIII,

(commercially available, e.g. from TCI, USA), especially under Stillecoupling and analogous/modified conditions; especially in an appropriatesolvent, especially a di-lower alkyl-lower alkanoyl amide, preferablydimethyl formamide or -acetamide; the compound of formula VIIIpreferably being in slight molar excess over the compound of the formulaVII, e.g. in the 1.1- to 5-fold, especially in the 1.5 to 2.5-foldexcess, for example in 2.1-fold excess; in the presence of catalyticamounts of AsPh₃ (especially about 0.4 equivalents), PdCl₂(MeCN)₂(especially about 0.2 equivalents) and Cul (especially about 0.1equivalents); at elevated temperatures, e.g. in the range of 50 to 90°C., preferably about 80 to about 85° C. For further reaction conditionssee the detailed description under process variant (a) (“Reaction a)”)above for the synthesis of a compound of the formula I. The reactionconditions can be optimized for the particular substrates in accordancewith the know-how of the person havin skill in the art.

The invention also relates to the inverted method wherein instead of thecompound of the formula VII an analogue is used where instead of theSn(R)₃ moiety a iodine is present and instead of the compound of theformula VIII an analogue is used that has the moiety Sn(R)₃ instead ofthe iodine. The reaction conditions are then analogous to those undermethod a) presented above for the synthesis of a compound of the formulaI.

The invention also relates to a method of synthesis for epothilones Eand especially F of the formula IX,

wherein Q is hydrogen (epothilone E) or methyl (epothilone F), which ischaracterized in that a compound of the formula X

is epoxidized in the presence of a peroxide to the compound of theformula IX, preferably by using standard reaction conditions forepoxidation, more preferably by epoxidation in the presence of H₂O₂, abase, especially KHCO₃, acetonitrile and an appropriate solvent,especially an alcohol, e.g. methanol, at temperatures preferably in therange of 0 to 50° C., especially about 25° C. (in-situ-formation ifmethylperoxycarboximidic acid); or in the presence of(+)-diethyl-D-tartrate and titanium isopropoxide, then t-butylhydroperoxide in an appropriate solvent, e.g. methylenchloride andoptionally decane at low temperatures, e.g. −78 to 0° C., especiallyabout −40° C.

These reactions have inter alia the advantage to provide the finalproducts in high yield and good isomeric purity.

Pharmaceutical Preparations

The present invention also relates to the use of a compound of theformula I for the manufacture of a pharmaceutical formulation for useagainst a proliferative disease as defined above; or to a pharmaceuticalformulation for the treatment of said proliferative disease comprising acompound of the invention and a pharmaceutically acceptable carrier.

The compounds of the formula I are called active ingredient hereinafter.

The invention relates also to pharmaceutical compositions comprising anactive ingredient as defined above, for the treatment of a proliferativedisease, especially as defined above, and to the preparation ofpharmaceutical preparations for said treatment.

The invention relates also to a pharmaceutical composition that issuitable for administration to a warm-blooded animal, especially ahuman, for the treatment of a proliferative disease as definedhereinbefore, comprising an amount of an active ingredient, which iseffective for the treatment of said proliferative disease, together withat least one pharmaceutically acceptable carrier. The pharmaceuticalcompositions according to the invention are those for enteral, such asnasal, rectal or oral, or preferably parenteral, such as intramuscularor intravenous, administration to a warm-blooded animal (human oranimal), that comprise an effective dose of the pharmacologically activeingredient, alone or together with a significant amount of apharmaceutically acceptable carrier. The dose of the active ingredientdepends on the species of warm-blooded animal, the body weight, the ageand the individual condition, individual pharmacokinetic data, thedisease to be treated and the mode of administration; preferably, thedose is one of the preferred doses as defined below, being accommodatedappropriately where pediatric treatment is intended.

The pharmaceutical compositions comprise from about 0.00002 to about95%, especially (e.g. in the case of infusion dilutions that are readyfor use) of 0.0001 to 0.02%, or (for example in case of infusionconcentrates) from about 0.1% to about 95%, preferably from about 20% toabout 90%, active ingredient (weight by weight, in each case).Pharmaceutical compositions according to the invention may be, forexample, in unit dose form, such as in the form of ampoules, vials,suppositories, dragees, tablets or capsules.

The pharmaceutical compositions of the present invention are prepared ina manner known per se, for example by means of conventional dissolving,lyophilizing, mixing, granulating or confectioning processes.

Solutions of the active ingredient, and also suspensions, and especiallyisotonic aqueous solutions or suspensions, are preferably used, it beingpossible, for example in the case of lyophilized compositions thatcomprise the active ingredient alone or together with a pharmaceuticallyacceptable carrier, for example mannitol, for such solutions orsuspensions to be produced prior to use. The pharmaceutical compositionsmay be sterilized and/or may comprise excipients, for examplepreservatives, stabilizers, wetting and/or emulsifying agents,solubilizers, salts for regulating the osmotic pressure and/or buffers,and are prepared in a manner known per se, for example by means ofconventional dissolving or lyophilizing processes. The said solutions orsuspensions may comprise viscosity-increasing substances, such as sodiumcarboxymethylcellulose, carboxymethylcellulose, dextran,polyvinylpyrrolidone or gelatin.

Suspensions in oil comprise as the oil component the vegetable,synthetic or semi-synthetic oils customary for injection purposes. Theremay be mentioned as such especially liquid fatty acid esters thatcontain as the acid component a long-chained fatty acid having from 8 to22, especially from 12 to 22, carbon atoms, for example lauric acid,tridecylic acid, myristic acid, pentadecylic acid, palmitic acid,margaric acid, stearic acid, arachidic acid, behenic acid orcorresponding unsaturated acids, for example oleic acid, elaidic acid,erucic acid, brasidic acid or linoleic acid, if desired with theaddition of anti-oxidants, for example vitamin E, betacarotene or3,5-di-tert-butyl-4-hydroxytoluene. The alcohol component of those fattyacid esters has a maximum of 6 carbon atoms and is a mono- orpolyhydroxy, for example a mono-, di- or tri-hydroxy, alcohol, forexample methanol, ethanol, propanol, butanol or pentanol or the isomersthereof, but especially glycol and glycerol.

The injection or infusion compositions are prepared in customary mannerunder sterile conditions; the same applies also to introducing thecompositions into ampoules or vials and sealing the containers.

Preferred is an infusion formulation comprising an active ingredient anda pharmaceutically acceptable organic solvent.

The pharmaceutically acceptable organic solvent used in a formulationaccording to the invention may be chosen from any such organic solventknown in the art. Preferably the solvent is selected from alcohol, e.g.absolute ethanol or ethanol/water mixtures, more preferably 70% ethanol,polyethylene glycol 300, polyethylene glycol 400, polypropylene glycolor N-methylpyrrolidone, most preferably polypropylene glycol or 70%ethanol or polyethylene glycol 300.

The active ingredient may preferably be present in the formulation in aconcentration of about 0.01 to about 100 mg/ml, more preferably about0.1 to about 100 mg/ml, still more preferably about 1 to about 10 mg/ml(especially in infusion concentrates).

The active ingredient may be used as pure substances or as a mixturewith another active ingredient. When used in its pure form it ispreferable to employ a concentration of active ingredient of 0.01 to100, more preferably 0.05 to 50, still more preferably 1 to 10 mg/ml(this number makes reference especially to an infusion concentrate that,before treatment, is diluted accordingly, see below).

Such formulations are conveniently stored in vials or ampoules.Typically the vials or ampoules are made from glass, e.g. borosilicateor soda-lime glass. The vials or ampoules may be of any volumeconventional in the art, preferably they are of a size sufficient toaccommodate 0.5 to 5 ml of formulation. The formulation is stable forperiods of storage of up to 12 to 24 months at temperatures of at least2 to 8° C.

Formulations must be diluted in an aqueous medium suitable forintravenous administration before the formulation of the activeingredient can be administered to a patient.

The infusion solution preferably must have the same or essentially thesame osmotic pressure as body fluid. Accordingly, the aqueous mediumpreferably contains an isotonic agent which has the effect of renderingthe osmotic pressure of the infusion solution the same or essentiallythe same as body fluid.

The isotonic agent may be selected from any of those known in the art,e.g. mannitol, dextrose, glucose and sodium chloride. Preferably theisotonic agent is glucose or sodium chloride. The isotonic agents may beused in amounts which impart to the infusion solution the same oressentially the same osmotic pressure as body fluid. The precisequantities needed can be determined by routine experimentation and willdepend upon the composition of the infusion solution and the nature ofthe isotonic agent. Selection of a particular isotonic agent is madehaving regard to the properties of the active agent.

The concentration of isotonic agent in the aqueous medium will dependupon the nature of the particular isotonic agent used. When glucose isused it is preferably used in a concentration of from 1 to 5% w/v, moreparticularly 5% w/v. When the isotonic agent is sodium chloride it ispreferably employed in amounts of up to 1% w/v, in particular 0.9% w/v.

The infusion formulation may be diluted with the aqueous medium. Theamount of aqueous medium employed as a diluent is chosen according tothe desired concentration of active ingredient in the infusion solution.Preferably the infusion solution is made by mixing a vial or ampoule ofinfusion concentrate afore-mentioned with an aqueous medium, making thevolume up to between 20 ml and 200 ml, preferably between about 50 andabout 100 ml, with the aqueous medium.

Infusion solutions may contain other excipients commonly employed informulations to be administered intravenously. Excipients includeantioxidants. Infusion solutions may be prepared by mixing an ampoule orvial of the formulation with the aqueous medium, e.g. a 5% w/v glucosesolution in WFI or especially 0.9% sodium chloride solution in asuitable container, e.g. an infusion bag or bottle. The infusionsolution, once formed, is preferably used immediately or within a shorttime of being formed, e.g. within 6 hours. Containers for holding theinfusion solutions may be chosen from any conventional container whichis nonreactive with the infusion solution. Glass containers made fromthose glass types afore-mentioned are suitable although it may bepreferred to use plastics containers, e.g. plastics infusion bags.

The invention also relates to a method of treatment of a warm-bloodedanimal, especially a human, that is in need of such treatment,especially of treatment of a proliferative disease, comprisingadministering a compound of the formula I, or a pharmaceuticallyacceptable salt thereof, to said warm-blooded animal, especially ahuman, in an amount that is sufficient for said treatment, especiallyeffective against said proliferative disease.

Dosage forms may be conveniently administered intravenously in a dosageof from 0.01 mg up to 100 mg/m² of active ingredient, preferably from0.1 to 20 mg/m² of active ingredient. The exact dosage required and theduration of administration will depend upon the seriousness of thecondition, the condition of the patient and the rate of administration.The dose may be administered daily or preferably with intervals of somedays or weeks, for example weekly or every 3 weeks. As the dose may bedelivered intravenously, the dose received and the blood concentrationcan be determined accurately on the basis of known in vivo and in vitrotechniques.

Pharmaceutical compositions for oral administration can be obtained bycombining the active ingredient with solid carriers, if desiredgranulating a resulting mixture, and processing the mixture, if desiredor necessary, after the addition of appropriate excipients, intotablets, dragee cores or capsules. It is also possible for them to beincorporated into plastics carriers that allow the active ingredients todiffuse or be released in measured amounts.

The compounds of the invention can be used alone or in combination withother pharmaceutically active substances, e.g. with otherchemotherapeutics, such as classical cytostatics. In the case ofcombinations with an other chemotherapeutic, a fixed combination of twoor more components or two or more independent formulations (e.g. in akit of part) are prepared as described above, or the otherchemotherapeutics are used in standard formulations that are marketedand known to the person of skill in the art, and the compound of thepresent invention and any other chemotherapeutic are administered at aninterval that allows for a common, additional or preferably synergisticeffect for tumor treatment.

The following examples are intended to illustrate the present inventionwithout being intended to limit the scope of the invention.

EXAMPLE 1

5-Methylpyridine Analogue D of Epothilone B

To a solution of B (20 mg, 0.035 mmol) in degassed dimethyl formamide(=DMF; 350 μl, 0.1 M) is added C (17 mg, 0.077 mmol, 2.1 equivalents)followed by AsPh₃ (4 mg, 0.4 equivalents), PdCl₂(MeCN)₂ (2 mg, 0.2equivalents) and Cul (1 mg, 0.1 equivalents) and the resulting slurry isplaced in an oil bath at 80-85° C. for 25 minutes. The reaction mixtureis then cooled to room temperature and the DMF removed by distillation.The residue is taken up in ethyl acetate, filtered through a small plugof silica, and eluted with hexanelethyl acetate (1/1, v/v). The solutionis then concentrated in vacuo and purified by preparative TLC(hexane/ethyl acetate 1/2) to give compound D: MS (Electrospray):expected: (M+H)⁺=502, found: 502. ¹H-NMR (600 MHz, CDCl₃): 8.37 (s, 1H,pyridine H); 7.50 (d, I=7.5 Hz, 1H, pyridine H); 7.19 (d, I=7.5 Hz, 1H,pyridine H), 6.59 (s, 1H, ═CH pyridine).

Starting Materials

To a solution of 7002 (see Scheme 11 below) (55 mg, 0.1 mmol) in toluene(1 ml, 0.1 M), Hünig's base is added (4 μl, 0.2 equivalents.), as wellas Pd(PPh₃)₄ (12 mg, 0.1 equivalents) and (Me₆)Sn₂ (91 μl, 5equivalents). This solution is then warmed to 80-85° C. for 15 minutes,and then cooled to room temperature. The yellow solution is thenfiltered through a small plug of silica, and eluted with hexane/ethylacetate (1/1, v/v). The solution is then concentrated on vacuo and theresidue is purified by flash chromatography (hexane/diethyl-ether 1/1 tohexane/ethyl acetate 1/1) to give B (20.2 mg, 34.4%) as a waxy solid.Data for B: HRMS (FAB Cs): Expected: M+Cs=703/705/707.1276, found:703/705/707.1402. ¹H-NMR (600 MHz, CDCl₃): 5.88 (t, J_(H-Sn)=40.6 Hz,1H, ═CH—SnMe₃) and 0.18 (t, J_(H-Sn)=40.6 Hz, 9H, SnMe₃).

EXAMPLE 2

In Analogy to Example 1 the Following Examples are Prepared

Starting Compound R₁ Physical Data Material Example 2 A)

Yield = 40%; MS (FAB) Expected (M + Cs): 620.1988, found; 620.2010;¹H-NMR (500 MHz, CDCl₃): 8.53 (d, J = 4.4 Hz, 1H, pyridine H); 7.72 (t,J = 7.4 Hz, 1H, pyridine H); 7.31 (d, J = 7.7 Hz, 1H, pyridine H); 7.17(t, J = 6.6 Hz, 1H, pyridine H); 6.65 (s, 1H, ═CH pyridine).

Example 2 B)

Yield = 14%; MS (FAB): expected (M + Cs): 634.2145; found 634.2122;¹H-NMR (500 MHz, CDCl₃): 8.36 (d, J = 4.4 Hz, 1H, pyridine H); 7.53 (d,J = 7.4 Hz, 1H, pyridine H); 7.13 (dd, J = 4.4, 7.4 Hz, 1H, pyridine H);6.64 (s, 1H, ═CH pyridine).

Example 2 C)

Yield = 20%; MS (FAB) expected (M + Cs): 634.2145; found 634.2124;¹H-NMR (500 MHz, CDCl₃): 8.38 (d, J = 4.8 Hz, 1H, pyridine H); 7.11 (s,1H, pyridine H); 6.98 (d, J = 4.8 Hz, 1H, pyridine H); 6.59 (s, 1H, ═CHpyridine).

EXAMPLE 3

Total Synthesis of Epothilone E and Related Side-Chain Modified Analogsvia a Stille Coupling Based Strategy

The first total synthesis of epothilone E (3) is accomplished by astrategy in which the key step is a Stille coupling (Stille et al.Angew. Chem. Int. Ed. Engl. 1986, 25, 508-524; Farina et al. J. Org.React. 1997, 50, 1-65) between vinyl iodide 7 and the thiazole moiety(8h, Scheme 2a). The macrolactone core fragment 7, which is prepared viaring-closing olefin metathesis (RCM), is subsequently used to provideconvenient and flexible access to a variety of side-chain modifiedepothilone analogs (9) for biological evaluation (Scheme 2b). The RCMreaction used to access 7 also provides trans-macrolactone (11, Scheme2b) which serves as an alternative template for the Stille couplingprocess and provides an additional array of analogs 10.

The chemical synthesis of the requisite vinyl iodides 7 and 11 isdelineated in PCT application WO 98/25929.

The stannane coupling partners used in the Stille reaction are shown inScheme 3.

Stille coupling: Procedure A: 2.0 equiv. of 8, 5-10 mol % Pd(PPh₃)₄,toluene, 90-100° C., 15-40 min, 39-88%; procedure B: 2.0-2.2 equiv. of8, 20-30 mol % Pd(MeCN)₂Cl₂, DMF, 25° C., 12-33 h, 49-94%.

The coupling partners 8b, 8d, 8h and 8j and additional stannanes 8p-rare prepared from readily accessible 2,4-dibromothiazole (20) viamonobromides 21 as outlined in Schemes 4 and 5.

Preparation of A) stannanes 8b, 8d and 8p. Reagents and conditions: (b)3.0 equiv. of NaSMe, EtOH, 25° C., 2 h, 92%; (d) 13 equiv. NaOH, EtOH,25° C., 30 h, 91%; (e) 13 equiv. NaOH, MeOH, 25° C., 16 h, 82%; (f) 5-10equiv. of Me₃SnSnMe₃, 5-10 mol % of Pd(PPh₃)₄, toluene, 80-100° C.,0.5-3 h, 81-100%; (g) 1.1 equiv. of n-BuLi, 1.2 equiv. of n-Bu₃SnCl, −78to 25° C., 30 min, 98%;

Preparation of stannanes 8h, 8j and 8q-s. Reagents and conditions: (a)1.05 equiv. n-Bu₃SnCH═CH₂, toluene, 100° C., 21 h, 83%; (b) 1.1-1.2equiv. of n-BuLi, 1.2-1.25 equiv. of n-Bu₃SnCl, −78 to 25° C., 1 h,28-85%; (c) H₂, 0.15 equiv. of PtO₂, EtOH, 25° C., 4 h; 84%; (d) 1.2equiv. of n-BuLi, 2.0 equiv. of DMF, −78 to 25° C., 2 h; (e) 1.9 equiv.of NaBH₄, MeOH, 25° C., 30 min, 63% for two steps; (f) 1.3 equiv. ofTBSCI, 2.0 equiv. of imidazole, CH₂Cl₂, 25° C., 0.5 h, 96%; (h) 1.2equiv. of TBAF, THF, 25° C., 20 min, 95%; (j) 1.1 equiv. of DAST,CH₂Cl₂, −78 to 25° C., 10 min, 57%. DAST=diethylamino sulfurtrifluoride.

Sulfide 21b is obtained in 92% yield by replacing the 2-bromosubstituent of 20 with the thiomethyl moiety using sodium thiomethoxide(EtOH, 25° C.). The ethoxy and methoxy thiazoles 21d and 21p areprepared by treating dibromide 20 with NaOH in ethanol and methanol,respectively. Bromides (21b, 21d and 21p) are then transformed to thedesired trimethylstannanes (8b, p) with hexamethylditin under palladiumcatalyzed conditions [Pd(PPh₃)₄, toluene, 80-100° C.], whereastri-n-butylstannane 8d is obtained from ethoxy-bromide 21d byhalogen-metal exchange (n-BuLi, Et₂O, −78° C.) and subsequent trappingwith tri-n-butyltin chloride in 98% yield.

The synthesis of stannanes (8h, 8j 8q-r) is also achieved from thecommon precursor 20 (Scheme 5). Thus, palladium catalyzed alkenylation[n-Bu₃SnCH═CH₂, Pd(PPh₃)₄, toluene, 100° C.] of 2,4-dibromothiazole 20affords monobromide 21q, which undergoes halogen-metal exchange (n-BuLi,Et₂O, −78° C.) and subsequent quenching with tri-n-butyltin chloride tofurnish the desired stannane 8q. Reduction of the intermediate vinylbromide 21q (H₂, PtO₂, EtOH, 25° C.) provides access to ethyl thiazole21r, which is converted into stannane 8r in an identical manner to thatdescribed for 8q. The synthesis of stannanes 8h and 8j is achieved viathe key hydroxymethyl thiazole 21h.

As shown in Scheme 5, this alcohol is, itself, obtained from dibromide20 in a two-step process involving lithiation (n-BuLi, Et₂O, −78° C.)and subsequent quenching with DMF to give intermediate aldehyde 22,which is then reduced (NaBH₄, MeOH, 25° C.) to furnish the desiredalcohol 21h in 63% overall yield. Conversion of 21h into stannane 8hrequires a three-step sequence involving protection of the hydroxylgroup (TBSCI, imidazole, CH₂Cl₂, 96%), stannylation (i. n-BuLi, Et₂O,−78° C.; ii. n-Bu₃SnCl, 85%) and subsequent deprotection (TBAF, THF, 25°C., 95%). Fluorination of the resulting stannane 8h (DAST, CH₂Cl₂, −78°C.) provides direct access to stannane 8j in 57% yield.

With the necessary components in hand, the critical Stille couplings areinvestigated. Two alternative sets of reaction conditions prove adequate(Scheme 3). Procedure A involves heating a toluene solution of thedesired vinyl iodide (7 or 11) with the appropriate stannane 8 in thepresence of catalytic amounts of Pd(PPh₃)₄ at 80-100° C. for between 15and 40 min. This protocol is used to couple stannanes 8b and 8h. Theremaining stannanes, 8d and 8j are coupled using an alternative, mildermethod, procedure B, in which a mixture of vinyl iodide (7 or 11) andstannane 8 in DMF is treated with PdCl₂(MeCN)₂ at 25° C.

The coupling of vinyl iodide 7 and stannane 8h provides macrolactone 18hwhich serves as the precursor to the natural epothilone E (3) (Scheme6a).

Preparation of epoxide 3. Reagents and conditions: (a) 30 equiv. ofH₂O₂, 60 equiv. of CH₃CN, 10 equiv. of KHCO₃, MeOH, 25° C., 6 h, 66%(based on 50% conversion).

The total synthesis is completed by epoxidation with in situ generatedmethylperoxycarboximidic acid (H₂O₂, KHCO₃, MeCN, MeOH, 25° C.;Chaudhuri et al. J. J. Org. Chem. 1982, 47, 5196-5198) furnishingepothilone E (3) (66% based on 50% conversion), which exhibits identicalphysical characteristics (¹H NMR, [α]_(D)) to those published in theart.

The Stille coupling approach is then extended to provide easy access toa variety of side-chain modified analogs of epothilone B (2), both atC-26 and the side chain. The retrosynthetic analysis of epothiloneanalogs possessing these dual modifications is shown in Scheme 6b andrequires the preparation of the crucial vinyl iodide core fragment 24. Amacrolactonization strategy similar to that used in our synthesis ofepothilone Band a variety of epothilone analogs is thought to be mostsuitable for this task.

Illustration of a retrosynthetic analysis of epothilone analogspossessing modified C26 and side-chain moieties.

The synthesis begins from the vinyl iodide 13 (Scheme 7) used in thepreparation of epothilone E and related analogs (Scheme 3).

Stereoselective synthesis of aldehyde 35. Reagents and conditions: (a)1.7 equiv. TBSCI, 2.8 equiv. imidazole, DMF, 0 to 25° C., 7 h, 84%; (b)i. 1.0 mol % OsO₄, 1.1 equiv. NMO, THF:t-BuOH:H₂O (5:5:1), 0 to 25° C.,13 h, 89%; ii. 6.0 equiv. NaIO₄, MeOH:H₂O (2:1), 0° C., 30 min, 92%; (c)2.4 equiv. 27, benzene, reflux, 1.2 h, 98%; (d) 3.0 equiv. DIBAL, THF,−78° C., 2.5 h, 100%. (e) 1.4 equiv. of TrCl, 1.7 equiv. of 4-DMAP, DMF,80° C., 21 h, 95%; (f) 1.4 equiv. of 9-BBN, THF, 0° C., 9 h; then 3 Naqueous NaOH and 30% H₂O₂, 0° C., 1 h, 95%; (g) 2.6 equiv. of I₂, 5.0equiv. of imidazole, 2.5 equiv. of Ph₃P, Et₂O:MeCN (3:1), 0° C., 45 min,97%; (h) 1.3 equiv. of the SAMP hydrazone of propionaldehyde, 1.4 equiv.of LDA, THF, 0° C., 16 h; then −100° C. and add 1.0 equiv. of 32 in THF,−100 to −20° C., 20 h, 71%; (i) 2.5 equiv. of MMPP, MeOH:phosphatebuffer pH 7 (1:1), 0° C., 3.5 h, 89%; (j) 3.0 equiv. of DIBAL, toluene,−78° C., 1 h, 88%. 9-BBN=9-borabicyclo[3.3.1]nonane;DIBAL=diisobutylaiuminium hydride; 4-DMAP=4-dimethylaminopyridine;LDA=lithium diisopropylamide; NMO=4-methylmorpholine N-oxide;SAMP=(S)-(−)-1-amino-2-(methoxymethyl)pyrrolidine;MMPP=monoperoxyphthalic acid, magnesium salt.

Protection of the allylic hydroxyl group (TBSCI, imidazole, DMF, 0 to25° C.) affords silyl ether 25 (84%) which is transformed into aldehyde26 by a two-step dihydroxylation-glycol-cleavage sequence (OsO₄, NMO,THF/t-BuOH/H₂O, 0 to 25° C.; then NaIO₄, MeOH/H₂O, 0° C., 82% for twosteps). A stereocontrolled Wittig reaction with the stabilized ylide 27(benzene, reflux; Marshall et al. J. Org. Chem. 1986, 51, 1735-1741;Bestmann et al. Angew. Chem. Int. Ed. Engl. 1965, 4, 645-660.) affordsester 28 as a single geometrical isomer in 98% yield. Reduction of thelatter compound (DIBAL, THF, −78° C.) affords alcohol 29, which isprotected as the triphenylmethyl (trityl) derivative 30 (TrCl, 4-DMAP,DMF, 70° C., 95%).

Elaboration of the terminal olefin is then achieved by selectivehydroboration-oxidation to give alcohol 31 (9-BBN, THF, 0° C.; thenNaOH, H₂O₂, 0° C.) which is transformed further into diiodide 32 (I₂,imidazole, Ph₃P, 0° C.) in 92% overall yield. Introduction of the C8stereocenter is then achieved using an Ender's alkylation protocol (SAMPhydrazone of propionaldehyde, LDA, THF, 0° C.; then −100° C. and add 32in THF; Enders et al. Asymmetric Synthesis 1984; Morrison, J. D., Ed.;Academic Press, Orlando, Vol 3, p. 275-339; we thank Prof. Enders for agenerous gift of SAMP) resulting in the formation of SAMP hydrazone 33in 71% yield. Conversion to nitrile 34 (MMPP, MeOH/phosphate buffer pH7, 0° C., 89%) and ensuing reduction (DIBAL, toluene, −78° C.) affordthe desired aldehyde 35 in 88% yield.

The transformation of aldehyde 35 into the desired epothilonemacrocyclic core 24 is summarized in Scheme 8.

Stereoselective synthesis of vinyl iodide 24. Reagents and conditions:(a) 1.45 equiv. of LDA, THF, −78° C., then 1.4 equiv. of 36 in THF, −78°C., 1.5 h then; −40° C., 0.5 h; then 1.0 equiv. of 35 in THF at −78° C.(66% combined yield, ca. 1.5:1 ratio of 37:38); (b) 3.2 equiv. ofTBSOTf, 4.3 equiv. of 2,6-lutidine, CH₂Cl₂, −20 to 0° C., 2.5 h, 90%;(c) HF.pyr. in pyridine, THF, 0° C., 3 h, 84%; (d) 2.0 equiv. of(COCl)₂, 4.0 equiv. of DMSO, 6.0 equiv. of Et₃N, CH₂Cl₂, −78 to 0° C.,1.5 h, 98%; (e) 5.0 equiv. of NaClO₂, 75 equiv. of 2-methyl-2-butene,2.5 equiv. of NaH₂PO₄, t-BuOH:H₂O (4.5:1), 25° C., 40 min, 100%; (f) 6.0equiv. of TBAF, THF, 0 to 25° C., 19 h, 95%; (g) 6.0 equiv. of Et₃N, 2.4equiv. of 2,4,6-trichlorobenzoylchloride, THF, 0° C., 1.5 h; then add toa solution of 2.2 equiv. of 4-DMAP in toluene (0.005 M based on 43), 75°C., 2.5 h, 84%; (h) 25% v/v HF.pyr. in THF 0 to 25° C., 15 h, 86%.TBAF=tetra n-butylammonium fluoride.

Aldol reaction of ketone 36, previously used in our synthesis ofepothilone B and related analogs (LDA, THF, −78 to −40° C.) and aldehyde35, affords alcohols 37 and 38 in 66% overall yield, with modestselectivity for the desired 6R,7S diastereoisomer (37). Separation andsilylation (TBSOTf, 2,6-lutidine, CH₂Cl₂, −20 to 0° C.) of the correctaldol product 37 provides tris-silyl ether 39 in 90% yield. Selectiveremoval of the primary silyl ether protecting group (HF.pyr. inpyridine/THF, 0° C.) affords alcohol 40 (84%), which is oxidized to acid42 via aldehyde 41 by a two-step procedure [Swern; then NaClO₂,2-methyl-2-butene, NaH₂PO₄, t-BuOH/H₂O, 25° C., 98% for two steps).Removal of the C15 silicon protecting group (TBAF, THF, 0 to 25° C.)provides hydroxy-acid 43 (95%) and lays the foundation for themacrolactonization process. This key step is achieved under Yamaguchiconditions (2,4,6-trichlorobenzoylchloride, Et₃N, THF; then add to asolution of 4-DMAP in toluene, 0.005M, 75° C.; Inanaga et al. Bull.Chem. Soc. Jpn. 1979, 52, 1989; Mulzer et al. Synthesis 1992, 215-228;Nicolaou et al. Chem. Eur. J. 1996, 2, 847-868) to give the protectedepothilone core 44 in 84% yield. Global deprotection (HF.pyr., THF, 0 to25° C., 86%) completes the synthesis of the key vinyl iodideintermediate 24.

With intermediate 24 in hand, the Stille coupling protocol is thenemployed to attach the desired heterocyclic moiety. The mild procedureB, employing PdCl₂(MeCN)₂ is originally thought to be the most practicaland efficient process and is utilized in the preparation of C26 hydroxyepothilones 45-48 (Scheme 9) from the vinyl iodide 24 and theappropriate stannanes 8 (see Schemes 4 and 5).

Synthesis of epothilone analogs 54-56 and 58, 59 and desoxyepothilones45-49 and 50-53. Reagents and conditions: (a) procedure A: 1.7 equiv. of8, 13 mol % Pd(PPh₃)₄, toluene, 100° C., 2 h, 15%; procedure B: 1.5-2.0equiv. of 8, 10-20 mol % Pd(MeCN)₂Cl₂, DMF, 25° C., 15-33 h, 41-56%; (b)1.05-1.4 equiv. of DAST, CH₂Cl₂, −78° C., 10 min, 26-58%; (c) 0.5 equiv.(+)-DET, 0.5 equiv. Ti(i-PrO)₄, 2.2 equiv. of t-BuOOH, −40° C., CH₂Cl₂,4Å molecular sieves, 1-2 h, 52-89%. DET=diethyl tartrate.

Unfortunately, these conditions are not suitable for the coupling of 24and vinyl stannane 8q (see Scheme 5). Recourse to the alternativeprocedure A provides access to the desired epothilone 49, albeit, inpoor yield.

The presence of the C26 hydroxy functionality provides a convenienthandle for further elaboration of the epothilone products. For example,the C26 alcohols 45-47 and 49 are treated with DAST (CH₂Cl₂, −78° C.) tofurnish fluorinated epothilone analogs 50-53 in moderate yields as shownin Scheme 9. Alternatively, asymmetric epoxidation of substrates 45 and46 under Katsuki-Sharpless conditions [(+)-DET, Ti(i-PrO)₄, t-BuOOH, 4 Åmolecular sieves, CH₂Cl₂, −40° C.; Katsuki, T.; Sharpless, K. B. J. Am.Chem. Soc. 1980, 102, 5976-5978] affords epothilones 54 and 55,respectively. Subsequent treatment with DAST (CH₂Cl₂, −78° C.) providesadditional analogs 58 and 59, again in moderate yield. At this juncture,a more efficient approach to epoxides such as 54 and 55 is envisaged inwhich asymmetric epoxidation of vinyl iodide 24 is achieved to give acommon intermediate, which then serves as a substrate for the Stillecoupling. Despite initial reservations concerning the compatibility ofthe epoxide functionality with the Stille conditions, the epoxide 57required for this approach is prepared from olefin 24 in 81% yield asdescribed for the synthesis of 45 and 46. To our pleasant surprise,application of the standard coupling procedure B, using stannane 8r,results in the successful preparation of epothilone analog 56 (73% yieldbased on 70% conversion).

The success of the Stille coupling strategy on substrates possessing anepoxide moiety indicates that epothilones 66-68 can be accessed from acommon intermediate 65 as outlined in Scheme 10.

Synthesis of C26-substituted epothilones 66-68. Reagents and conditions:(a) 15 equiv. of Et₃N, 8.0 equiv. TMSCl, DMF, 25° C., 12 h; (b) silicagel, CH₂Cl₂, 25° C., 12 h, 98% for two steps; (c) 3.0 equiv. of NMO, 10mol % TPAP, CH₂Cl₂, 25° C., 40 min, 90%; (d) 9.7 equiv. of Ph₃P⁺CH₃Br⁻mixture with NaNH₂), THF, −5° C., 65% (e) 25 equiv. of H₂NNH₂, 16 equiv.of H₂O₂, EtOH, 0°, 3 h; (f) HF.pyr. pyridine in THF, 0 to 25° C., 2 h,75% for two step; (g) 1.7-2.3 equiv. of 8, 0.2-0.3 mol % Pd(MeCN)₂Cl₂,DMF, 25° C., 15-23 h, 52-79%. TPAP=tetrapropylammonium perruthenate.

Preparation of the desired template (65) is achieved by a five-stepsequence, which starts with global protection of triol 57 (TMSCl, Et₃N,DMF, 25° C.). Selective deprotection, using silica gel (CH₂Cl₂, 25° C.,98% for two steps), reveals the C26 primary hydroxyl functionality whichis then oxidized (TPAP, NMO, 4 Å molecular sieves, CH₂Cl₂, 25° C.) tofurnish aldehyde 62 in 90% yeild. Methylenation using methyltriphenylphosphonium bromide (Schlosser's “instant ylid” mix, THF, −5°C.; Schlosser, M.; Schaub, B. Chimia 1982, 36, 3965) furnishes olefin 63(65%) which undergoes reduction with in situ generated diimide (H₂NNH₂,H₂O₂, EtOH, 0° C.) to give intermediate 64. Deprotection of theremaining silyl ethers (HF.pyridine (=pyr.). in pyridine/THF, 0° C.)affords the desired vinyl iodide 65 in 75% yield for two steps. TheStille coupling procedure B described above is then used to accessepothilones 66-68 in moderate yields (Scheme 10).

The chemistry described in this example relies on a Stille couplingapproach to construct a series of epothilone analogs with diversity atthe side-chain or at both the side-chain and C26 site from a commonmacrocyclic intermediate.

EXAMPLE 4

Formulae of Compounds According to the Invention

TABLE Formulae of compounds according to the invention: En- try Compound1

2 (Formula: see under entry 1) 18j: X = OH 3

4 (Formula: see under entry 3) 18b: X = SMe 5

6 (Formula: see under entry 5) 19j: X = F 7

8 (Formula: see under entry 7) 19b: X = SMe 9

10 (Formula: see under entry 9) 46: X = OMe 11 (Formula: see under entry9) 47: X = CH₂CH₃ 12 (Formula: see under entry 9) 48: X = CH₂OH 13(Formula: see under entry 9) 49: X = CH═CH₂ 14

15 (Formula: see under entry 14) 51: X = OMe 16 (Formula: see underentry 14) 52: X = CH═CH₂ 17 (Formula: see under entry 14) 53: X = CH₂CH₃18

19 (Formula: see under entry 18) 55: X = OMe 20 (Formula: see underentry 18) 56: X = CH₂CH₃ 21

22 (Formula: See under entry 21) 59: X = OMe 23

24 (Formula: see under entry 23) 67: X = OMe 25 (Formula: see underentry 23) 68: X = CH₂CH₃ 26 epothilione A

EXAMPLE 5

Biological Results

In accordance with the methods described above (inhibition of tubulindepolymerization by a compound of the formula I is measured using pigbrain microtubuli, comparison with 25 μM epothilone B; cellular assaysare analogous to those described above for KB-31 cells), the resultsgiven in the following table are obtained for the mentioned compounds offormula I:

HCT- KB-31^(b) KB-8511^(c) A549^(d) HCT-15^(e) 116^(e) Com- Tubulin^(a)IC50 IC50 IC50 IC50 IC50 pound (%) [nM] [nM] [nM] [nM] [nM] D (Exam-88.9 0.108 0.105 0.17 0.247 0.209 ple 1) Example 89.9 0.153 0.163 0.240.298 0.373 2C ^(a)Induction of tubulin polymerisation at 5 μMconcentration of test compound versus epothilone B at 25 μM (in %).^(b)epidermoid ^(c)epidermoid (P-gp overexpressing) ^(d)lung ^(e)colon

DU145^(f) PC-3M^(f) MCF-7^(g) MCF-7/ADR^(h) Compound IC50 [nM] IC50 [nM]IC50 [nM] IC50 [nM] D (Example 1) 0.252 0.361 0.114 0.853 Example 2B0.320 0.498 0.144 1.31 ^(f)prostate ^(g)breast ^(h)breast (multidrugresistant)

EXAMPLE 6

Further Compounds of the Formula I

In analogy to the methods described above and below, the compoundsfalling under formula I are prepared that have the following formulae:

EXAMPLE 6 (i)

EXAMPLE 6(ii)

EXAMPLE 6 (iii)

EXAMPLE 6 (iv)

EXAMPLE 6 (v)

EXAMPLE 6 (vi)

EXAMPLE 6 (vii)

EXAMPLE 6 (viii)

EXAMPLE 6 (ix)

EXAMPLE 6 (x)

EXAMPLE 6 (xi)

EXAMPLE 6

Pharmaceutical Formulation

Epothilone analogue D (example 1) or the epothilone analogue of Example2 C) (15 mg) is dissolved in 98-100% propylene glycol (1.0 ml). Thesolution is sterile filtered through a 0.22 microns pore size filter andcharged to 1 ml ampoules. The filled ampoules are used for storage andshipment. Prior to intravenous administration, the contents of anampoule are added to 250 to 1000 ml of a 5% glucose solution inwater-for-injection.

EXAMPLE 7

Use of Additional Stannanes to Synthesize Side Chain Modified EpothiloneAnalogs as Illustrated in Schemes 11 and 12

General Route to the synthesis of various side-chain modified epothiloneB analogs having pyridine and imidazole modifications.

a: as previously described (see Nicolaou et al. Tetrahedron 54,7127-7166 (1998)); b, d, e: conditions as previously described (seeabove or Nicolaou et al. Tetrahedron 54, 7127-7166); c: NaBH₃CN, HMPA,40-45° C.

Protecting groups are those known in the art, especially those describedin the standard references mentioned hereinabove, as well as the methodsof their introduction and removal mentioned in said standard references.

Preferably, in 7006 R* is H or methyl. in 7004, R is preferably methyl.

Use of the Stille coupling procedure to prepare a number of side chainmodified epothilone analogs from the common precursors 57, and 8h, 8x,8y and 8z is described in Scheme 11 and 12. Synthesis of vinyl iodide7002 is achieved from the previously reported C26-hydroxy compound andinvolves conversion of 57 to diiodide 7001 and subsequent reductionusing NaBH₃CN: Diiodide 7001 (1 equivalent; from 57) and sodiumcyanoborohydride (10 equivalents) are dissolved in anhydrous HMPA (0.2M) and the resulting mixture heated at 45-50° C. for 48 h. After coolingto room temperature, water is added and the aqueous phase extracted fourtimes with ethyl acetate. The combined organic fractions are dried(Na₂SO₄) and passed through a short plug of silica gel to remove tracesof HMPA (eluting with 50% ethyl acetate in hexanes). Followingevaporation of solvents, the residue is purified by preparative thinlayer chromatography (eluting with 50% ethyl acetate in hexanes) toprovide pure vinyl iodide 7002 (84%). Coupling to the epothilone E sidechain is achieved and the coupling of a number of pyridines andimidazoles is accomplished via coupling of numerous alternative sidechains with the aromatic stannanes as shown in Schemes 11 and 12 usingthe standard methods outlined herein.

Illustration of some side chain modified epothilone analogs usingindicated aryl stannanes (ArSnR₃) from either the metathesis ormacrolactonization approach wherein R is n-butyl or methyl. Thestannanes are synthesized using standard conditions known in the art. Xis a radical selected from the group consisting of hydrogen; loweralkyl, especially methyl, ethyl, n-propyl, iso-propyl, n-butyl,iso-butyl, tert-butyl, n-pentyl, n-hexyl; —CH═CH₂; —C≡CH; —CH₂F; —CH₂Cl;—CH₂—OH; —CH₂—O—(C₁-C₆-alkyl), especially —CH₂—O—CH₃; and—CH₂—S—(C₁-C₆-alkyl), especially —CH₂—S—CH₃; and R is methyl or n-butyl.

Synthetic Protocols

General: All reactions are carried out under an argon atmosphere withdry, freshly distilled solvents under anhydrous conditions, unlessotherwise noted. Tetrahydrofuran (THF) and diethyl ether (ether) aredistilled from sodium-benzophenone, and dichloromethane (CH₂Cl₂),benzene (PhH), and toluene from calcium hydride. Anhydrous solvents arealso obtained by passing them through commercially available activatedalumina columns. Yields refer to chromatographically andspectroscopically (¹H NMR) homogeneous materials, unless otherwisestated. All solutions used in workup procedures are saturated unlessotherwise noted. All reagents are purchased at highest commercialquality and used without further purification unless otherwise stated.All reactions are monitored by thin-layer chromatography carried out on0.25 mm E. Merck silica gel plates (60F-254) using UV light asvisualizing agent and 7% ethanolic phosphomolybdic acid orp-anisaldehyde solution and heat as developing agents. E. Merck silicagel (60, particle size 0.040-0.063 mm) is used for flash columnchromatography. Preparative thin-layer chromatography separations arecarried out on 0.25, 0.50 or 1 mm E. Merck silica gel plates (60F-254).NMR spectra are recorded on Bruker DRX-600, AMX-500, AMX-400 or AC-250instruments and calibrated using residual undeuterated solvent as aninternal reference. The following abbreviations are used to explain themultiplicities: s, singlet; d, doublet; t, triplet; q, quartet; m,multiplet; band, several overlapping signals; b, broad. IR spectra arerecorded on a Perkin-Elmer 1600 series FT-IR spectrometer. Opticalrotations are recorded on a Perkin-Elmer 241 polarmeter. High resolutionmass spectra (HRMS) are recorded on a VG ZAB-ZSE mass spectrometer underfast atom bombardment (FAB) conditions.

cis-Macrolactone diol 7 as illustrated in Scheme 3. To a solution ofiodide 16 (305 mg, 0.491 mmol) in THF (8.2 mL, 0.06 M) at 25° C. isadded HF.pyr. (2.7 mL) and the resulting solution is stirred at the sametemperature for 27 h. The reaction is then quenched by careful additionto a mixture of saturated aqueous NaHCO₃ (100 mL) and EtOAc (100 mL),and the resulting two-phase mixture is stirred at 25° C. for 2 h. Theextracts are then separated and the organic layer is washed withsaturated aqueous NaHCO₃ (100 mL) and brine (100 mL), and then dried(MgSO₄). Purification by flash column chromatography (silica gel, 20 to50% EtOAc in hexanes) furnishes diol 7 (208 mg, 84%). R_(f)=0.21 (silicagel, 25% EtOAc in hexanes); [α]²² _(D) −53.1 (c 1.37, CHCl₃); IR (thinfilm) ν_(max) 3499 (br), 2930, 1732, 1688, 1469, 1379, 1259, 1149, 1093,1048, 1006, 732 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ6.43 (s, 1 H,ICH═C(CH₃)), 5.44 (ddd, J=10.5, 10.5, 4.5 Hz, 1 H, CH═CHCH₂), 5.34 (dd,J=9.5, 2.0 Hz, 1 H, CHOCO), 5.32 (ddd, J=10.5, 10.5, 5.5 Hz, 1 H,CH═CHCH₂), 4.07 (ddd, J=11.0, 6.0, 3.0 Hz, 1 H, (CH₃)₂CCH(OH)), 3.73(ddd, J=2.5, 2.5, 2.5 Hz, 1 H, CHOH(CHCH₃)), 3.10 (qd, J=7.0, 2.5 Hz, 1H, CH₃CH(C═O)), 2.84 (d, J=2.5 Hz, 1 H, CH(CH₃)CHOHCH(CH₃)), 2.66 (ddd,J=15.0, 9.5, 9.5 Hz, 1 H, ═CHCH₂CHO), 2.51 (dd, J=15.5, 11.0 Hz, 1 H,CH₂COO), 2.42 (dd, J=15.5, 3.0 Hz, 1 H, CH₂COO), 2.35 (d, J=6.0 Hz, 1 H,(CH₃)₂CHOH), 2.21-2.12 (m, 2 H), 2.05-1.97 (m, 1 H), 1.88 (s, 3 H,ICH═CCH₃), 1.76-1.70 (m, 1 H), 1.70-1.62 (m, 1 H), 1.32 (s, 3 H,C(CH₃)₂), 1.18 (d, J=7.0 Hz, 3 H, CH₃CH(C═O)), 1.10 (s, 3 H, C(CH₃)₂),1.35-1.05 (m, 3 H), 0.99 (d, J=7.0 Hz, 3 H, CH₃CHCH₂); HRMS (FAB),calcd. for C₂₂H₃₅IO₅ (M+Cs⁺) 639.0584, found 639.0557.

trans-Macrolactone diol 11 as illustrated in Scheme 3. A solution ofiodide 17 (194 mg, 0.313 mmol) in THF (5.2 mL, 0.06 M) is treated withHF.pyr. (1.7 mL) according to the procedure described for thepreparation of diol 7 to afford, after flash column chromatography(silica gel, 20 to 50% EtOAc in hexanes), diol 11 (134 mg, 85%).R_(f)=0.16 (silica gel, 25% EtOAc in hexanes); [α]²² _(D) −20.0 (c 1.15,CHCl₃); IR (film) ν_(max) 3478, 2930, 1732, 1693 cm⁻¹; ¹H NMR (500 MHz,CDCl₃) δ6.37 (d, J=1.5 Hz, 1 H, ICH═CCH₃), 5.35 (ddd, J=14.5, 7.0, 7.0Hz, 1 H, CH═CHCH₂), 5.24 (ddd, J=14.5, 7.0, 7.0 Hz, 1 H, CH═CHCH₂), 5.17(dd, J=6.5, 3.5 Hz, 1 H, CHOCO), 4.41 (dd, J=8.0, 3.5 Hz, 1 H,(CH₃)₂CCH(OTBS)), 3.85 (bs, 1 H, CHOH(CHCH₃)), 3.38 (bs, 1 H,CHOH(CHCH₃)), 3.18 (qd, J=7.0, 6.5 Hz, 1 H, CH₃CH(C═O)), 2.68-2.34 (m, 4H), 2.44 (s, 3 H, CH₃Ar), 2.19-2.11 (m, 1 H), 1.96 (s, 3 H, CH₃C═CH),1.99-1.93 (m, 1 H), 1.67-1.52 (m, 2 H), 1.48-1.42 (m, 1 H), 1.31-0.99(m, 2 H), 1.22 (d, J=7.0 Hz, 3 H, CH₃CH(C═O)), 1.14 (s, 3 H, C(CH₃)₂),1.09 (s, 3 H, C(CH₃)₂), 1.02 (d, J=7.0 Hz, 3 H, CH₃CHCH₂), 0.84 (s, 9 H,SiC(CH₃)₃(CH₃)₂), 0.08 (s, 3 H, SiC(CH₃)₃(CH₃)₂), −0.01 (s, 3 H,SiC(CH₃)₃(CH₃)₂); HRMS (FAB), calcd. for C₂₂H₃₅IO₅ (M+Cs⁺) 639.0584,found 639.0606.

2-Thiomethyl-4-bromothiazole 21b as illustrated in Scheme 4.2,4-Dibromothiazole 20 (82 mg, 0.34 mmol, 1.0 equiv.) is dissolved inethanol (2.3 mL, 0.15 M) and treated with sodium thiomethoxide (75 mg,1.02 mmol, 3.0 equiv.). The reaction mixture is stirred at 25° C. for 2h, upon which time completion of the reaction is established by ¹H NMR.The mixture is poured into water (5 mL) and extracted with ether (2×5mL). The combined organic fractions are dried (MgSO₄), the solventsevaporated and the residue purified by flash column chromatography(silica gel, 5% EtOAc in hexanes) to furnish2-thiomethyl-4-bromothiazole 21b (77 mg, 92%). R_(f)=0.58 (silica gel,10% EtOAc in hexanes); IR (film) ν_(max) 3118, 2926, 1459, 1430, 1388,1242, 1040, 966, 876, 818 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ7.07 (s, 1 H,ArH), 2.69 (s, 3 H, SCH₃); GC/MS (EI), calcd. for C₄H₄BrNS₂ (M⁺)209/211, found 209/211.

2-Ethoxy-4-bromothiazole 21d as illustrated in Scheme 4. To a solutionof 2,4-dibromothiazole 20 (58 mg, 0.239 mmol, 1.0 equiv.) in EtOH (2.4mL, 0.1 M) is added NaOH (122 mg, 3.05 mmol, 12.8 equiv.) and theresulting solution is stirred at 25° C. until TLC indicates thedisappearance of dibromide (ca. 30 h). The resulting yellow solution isthen partitioned between ether (10 mL) and saturated aqueous NH₄Cl (10mL) and the layers are separated. The aqueous layer is extracted withether (10 mL) and the combined organic extracts are washed with brine(20 mL), dried (MgSO₄) and concentrated carefully under reducedpressure. Flash column chromatography (silica gel, 17% ether in hexanes)furnishes 2-ethoxy-4-bromothiazole 21d as a volatile oil (45 mg, 91%).R_(f)=0.58 (silica gel, 17% ether in hexanes); IR (film) ν_(max) 3125,2983, 2936, 2740, 1514, 1480, 1392, 1360, 1277, 1234, 1080, 1018, 897,823 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ6.57 (s, 1 H, ArH), 4.48 (q, J=7.0Hz, 2 H, CH₃CH₂), 1.43 (t, J=7.0 Hz, 3 H, CH₃CH₂); GC/MS (EI), calcd.for C₄H₄BrNSO (M⁺) 193/195, found 193/195.

2-Methoxy-4-bromothiazole 21p as illustrated in Scheme 4. To a solutionof 2,4-dibromothiazole 20 (253 mg, 1.04 mmol, 1.0 equiv.) in MeOH (10.5mL, 0.1 M) is added NaOH (555 mg, 13.9 mmol, 13.3 equiv.) and theresulting solution is stirred at 25° C. until TLC indicates thedisappearance of dibromide (ca. 16 h). The resulting yellow solution isthen partitioned between ether (10 mL) and saturated aqueous NH₄Cl (10mL) and the layers are separated. The aqueous phase is extracted withether (10 mL) and the combined organic extracts are dried (MgSO₄) andconcentrated carefully under reduced pressure. Flash columnchromatography (silica gel, 10% ether in hexanes) furnishes2-methoxy-4-bromothiazole 21p as a volatile oil (138 mg, 82%).R_(f)=0.56 (silica gel, 17% ether in hexanes); IR (film) ν_(max) 3125,2952, 2752, 1524, 1520, 1481, 1417, 1277, 1238, 1081, 982, 884, 819cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ6.58 (s, 1 H, ArH), 4.09 (q, 3 H, CH₃);GC/MS (EI), calcd. for C₅H₆BrNSO (M⁺) 207/209, found 207/209.

2-Hydroxymethyl-4-bromothiazole 21h as illustrated in Scheme 4. To asolution of 2,4-dibromothiazole 20 (50 mg, 0.206 mmol, 1.0 equiv.) inanhydrous ether (2.0 mL, 0.1 M) at −78° C., is added n-BuLi (154 μL, 1.6M in hexanes, 0.247 mmol, 1.2 equiv.), and the resulting solution isstirred at the same temperature for 30 min. DMF (32 μL, 0.412 mmol, 2.0equiv.) is then added at −78° C. and, after being stirred at −78° C. for30 min, the reaction mixture is slowly warmed up to 25° C. over a periodof 2 h. Hexane (2.0 mL) is added and the resulting mixture is passedthrough a short silica gel cake eluting with 30% EtOAc in hexanes. Thesolvents are evaporated to give the crude aldehyde 22 (50 mg), which isused directly in the next step.

To a solution of aldehyde 22 (50 mg) in methanol (2.0 mL) at 25° C., isadded sodium borohydride (15 mg, 0.397 mmol, 1.9 equiv.), and theresulting mixture is stirred at the same temperature for 30 min. EtOAc(1.0 mL) and hexane (2.0 mL) are added, and the mixture is passedthrough a short silica gel cake eluting with EtOAc. The solvents arethen evaporated and the crude product is purified by flash columnchromatography (silica gel, 20 to 50% EtOAc in hexanes) to furnish2-hydroxymethyl-4-bromothiazole 21h (25 mg, 63% over two steps).R_(f)=0.16 (silica gel, 18% EtOAc in hexanes); IR (film) ν_(max) 3288,3122, 2922, 2855, 1486, 1447, 1345, 1250, 1183, 1085, 1059, 967, 893cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ7.20 (s, 1 H, ArH), 4.93 (s, 2 H, CH₂);HRMS (FAB), calcd. for C₄H₄BrNOS (M+H⁺) 193.9275, found 193.9283.

2-(tert-Butyidimethylsilyloxymethyl)-4-bromothiazole 21s as illustratedin Scheme 5. To a solution of alcohol 21h (59 mg, 0.304 mmol, 1.0equiv.) in CH₂Cl₂ (1.0 mL, 0.3 M) is added imidazole (62 mg, 0.608 mmol,2.0 equiv.), followed by tert-butyldimethylchlorosilane (69 mg, 0.456mmol, 1.3 equiv.) at 25° C. After 30 min at 25° C., the reaction mixtureis quenched with methanol (100 mL) and then passed through silica geleluting with CH₂Cl₂. Evaporation of solvents gives the desired silylether 21s (90 mg, 96%). R_(f)=60 (silica gel, 10% EtOAc in hexanes); IR(film) ν_(max) 2943, 2858, 1489, 1465, 1355, 1254, 1193, 1108, 887, 841,780 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ7.16 (s, 1 H, ArH), 4.93 (s, 2 H,CH₂), 0,94 (s, 9 H, SiC(CH₃)₃(CH₃)₂), 0.12 (s, 6 H, SiC(CH₃)₃(CH₃)₂);HRMS (FAB), calcd. for C₁₀H₁₈BrNOSSi (M+H⁺) 308.0140, found 308.0151.

2-Vinyl-4-bromothiazole 21q as illustrated in Scheme 5. To a solution of2,4-dibromothiazole 20 (437 mg, 1.80 mmol, 1.0 equiv.) in toluene isadded tri-n-butyl(vinyl)tin (552 μL, 1.89 mmol, 1.05 equiv.) followed byPd(PPh₃)₄ (208 mg, 0.180 mmol, 0.1 equiv.) and the resuiting mixture isheated at 100° C. After 21 h, the mixture is cooled and purifieddirectly by flash column chromatography (silica gel, 0 to 9% ether inhexanes) to afford 2-vinyl-4-bromothiazole 21q as an oil (285 mg, 83%).R_(f)=0.50 (silica gel, 17% ether in hexanes); IR (film) ν_(max) 3121,1470, 1259, 1226, 1124, 1082, 975, 926, 887, 833 cm⁻¹; ¹H NMR (500 MHz,CDCl₃) δ7.13 (s, 1 H, ArH), 6.86 (dd, J=17.5,11.0 Hz, 1 H, CH═CH₂), 6.09(d, J=17.5 Hz, 1 H, CHCH₂), 5.59 (d, J=10.5 Hz, 1 H, CHCH₂); GC/MS (EI),calcd. for C₅H₄BrNS (M⁺) 189/191, found 189/191.

2-Ethyl-4-bromothiazole 21r as illustrated in Scheme 5. A solution of2-vinyl-4-bromothiazole 21q (279 mg, 1.47 mmol, 1.0 equiv.) in ethanol(15 mL, 0.1 M) is added PtO₂ (50 mg, 0.220 mmol, 0.15 equiv.) and theresulting mixture is stirred under an atmosphere of hydrogen at 25° C.for 4 h. Subsequent filtration through a short plug of silica gel,eluting with EtOAc, and careful concentration under reduced pressurefurnishes 2-ethyl-4-bromothiazole 21r (238 mg, 84%). R_(f)=0.63 (silicagel, CH₂Cl₂); IR (film) ν_(max) 3122, 2974, 2932, 1483, 1456, 1245,1181, 1090, 1040, 956, 884, 831 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ7.08 (s,1 H, ArH), 3.03 (q, J=7.5 Hz, 2 H, CH₂CH₃), 1.37 (t, J=7.5 Hz, 2 H,CH₂CH₃); GC/MS (EI), calcd. for C₅H₆BrNS (M⁺) 191/193, found 191/193.

2-Thiomethyl-4-trimethylstannylthiazole 8b as illustrated in Scheme 3.To a solution of bromothiazole 21b (51 mg, 0.24 mmol, 1.0 equiv.) indegassed toluene (4.9 mL, 0.1 M) is added hexamethylditin (498 μL, 2.4mmol, 10 equiv.) and Pd(PPh₃)₄ (14 mg, 0.012 mmol, 0.05 equiv.) and thereaction mixture is heated at 80° C. for 3 h. Then the reaction mixtureis cooled to 25° C. and affords, after flash column chromatography(silica gel, 5% Et₃N in hexanes), stannane 8b (71 mg, 100%). R_(f)=0.67(silica gel; pre-treated with Et₃N, 10% EtOAc); IR (film) ν_(max) 2981,2924, 1382, 1030, 772 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ7.25 (s, 1 H, ArH),2.70 (s, 3 H, SCH₃), 0.32 (s, 9 H, Sn(CH₃)₃); HRMS (FAB), calcd. forC₇H₁₃NS₂Sn (M+H⁺) 295.9588, found 295.9576.

2-Methoxy-4-trimethylstannylthiazole 8p as illustrated in Scheme 4. To asolution of bromothiazole 21p (147 mg, 0.758 mmol, 1.0 equiv.) indegassed toluene (7.6 mL, 0.1 M) is added hexamethyiditin (785 μL, 3.79mmol, 5.0 equiv.) and Pd(PPh₃)₄ (88 mg, 0.076 mmol, 0.1 equiv.) and thereaction mixture is heated at 100° C. for 30 min according to theprocedure described for the synthesis of stannane 8b to afford, afterflash column chromatography (silica gel, 5% Et₃N in hexanes), stannane8p (170 mg, 81%). R_(f)=0.49 (silica gel; pretreated with Et₃N, 17%ether in hexanes); IR (film) ν_(max) 2985, 2948, 2915, 1512, 1414, 1259,1234, 1219, 1087, 988 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ6.72 (s, 1 H, ArH),4.07 (s, 3 H, OCH₃), 0.32 (s, 9 H, Sn(CH₃)₃); HRMS (FAB), calcd. forC₇H₁₃NOSSn (M+H⁺) 279.9818, found 279.9810.

2-(tert-butyidimethylsilyloxymethyl)-4-tri-n-butylstannylthiazole 8s asillustrated in Scheme 5. To a solution of bromothiazole 21s (20 mg,0.065 mmol, 1.0 equiv.) in ether (1.0 mL, 0.07M) at −78° C., is addedn-BuLi (49 μL, 1.6 M in hexanes, 0.078 mmol, 1.2 equiv.) and theresulting mixture is stirred at −78° C. for 10 min. Tri-n-butyltinchloride (23 μL, 0.078 mmol, 1.2 equiv.) is then added, the solutionstirred at −78° C. for 10 min, and then slowly warmed to 25° C. over aperiod of 1 h. The reaction mixture is diluted with hexane (2.0 mL), andpassed through silica gel eluting with 20% EtOAc in hexanes. Flashcolumn chromatography (silica gel; pre-treated with Et₃N, 5% ether inhexanes) furnishes the desired stannane 8s (35 mg, 85%). R_(f)=0.36(silica gel, 5% EtOAc in hexanes); IR (film) ν_(max) 2955, 2928, 2856,1464, 1353, 1255, 1185, 1103, 1081, 1006, 841 cm⁻¹; ¹H NMR (500 MHz,C₆D₆) δ7.08 (s, 1 H, ArH), 4.98 (s, 2 H, CH₂), 1.75-1.57 (m, 6 H,CH₃CH₂), 1.44-1.31 (m, 6 H, CH₃CH₂CH₂), 1.26-1.09 (m, 6 H,CH₃CH₂CH₂CH₂), 0.94 (s, 9 H, SiC(CH₃)₃(CH₃)₂), 0.91 (t, J=7.0 Hz, 9 H,CH₃), −0.02 (s, 6 H, SiC(CH₃)₃(CH₃)₂); HRMS (FAB), calcd. forC₂₂H₄₅NOSSiSn (M+H⁺) 520.2093, found 520.2074.

2-Hydroxymethyl-4-tri-n-butylstannylthiazole 8h as illustrated in Scheme5. To a solution of silyl ether 8s (20 mg, 0.039 mmol, 1.0 equiv.) inTHF (1.0 mL, 0.04 M) is added TBAF (46 μL, 1.0 M in THF, 0.046 mmol, 1.2equiv.) and the reaction mixture is stirred at 25° C. for 20 min. Hexane(2.0 mL) is added, and the mixture passed through silica gel elutingwith EtOAc. Evaporation of solvents gives the desired alcohol 8h (15 mg,95%). R_(f)=0.09 (silica gel, 20% ether in hexanes); IR (film) ν_(max)3209, 2956, 2923, 2855, 1461, 1342, 1253, 1174, 1064, 962 cm⁻¹; ¹H NMR(500 MHz, CDCl₃) δ7.30 (m, 1 H, ArH), 4.99 (s, 2 H, CH₂), 3.64 (bs, 1 H,OH), 1.62-1.45 (m, 6 H, CH₃CH₂) 1.38-1.27 (m, 6 H, CH₃CH₂), 1.19-1.02(m, 6 H, CH₃CH₂CH₂CH₂), 0.88 (t, J=7.0 Hz, 9 H, CH₃); HRMS (FAB), calcd.for C₁₆H₃₁NOSSn (M+H⁺) 406.1228, found 406.1237.

2-Fluoromethyl-4-tri-n-butylstannylthiazole 8j as illustrated in Scheme5. To a solution of alcohol 8h (90 mg, 0.223 mmol, 1.0 equiv.) in CH₂Cl₂(2.2 mL, 0.1 M) at −78° C. is added DAST (32 μL, 0.242 mmol, 1.1 equiv.)and the solution is stirred at this temperature for 10 min. Afterquenching with saturated aqueous NaHCO₃ (2 mL) the mixture is allowed towarm up to 25° C., and then partitioned between CH₂Cl₂ (15 mL) andsaturated aqueous NaHCO₃ (15 mL). The layers are separated and theaqueous phase is extracted with CH₂Cl₂ (2×15 mL). The combined organicextracts are washed with brine (40 mL), dried (MgSO₄) and concentratedunder reduced pressure. Flash column chromatography (silica gel;pre-treated with Et₃N, 17% ether in hexanes) furnishes stannane 8j (52mg, 57%). R_(f)=0.59 (silica gel, 17% ether in hexanes); IR (film)ν_(max) 2956, 2925, 2870, 2863, 1464, 1376, 1358, 1184, 1084, 1023, 874,807 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) ε7.41 (s, 1 H, ArH), 5.69 (d, J=47.5Hz, 2 H, CH₂F), 1.58-1.52 (m, 6 H, (CH₃CH₂(CH₂)₂)₃Sn), 1.36-1.29 (m, 6H, (CH₃CH₂CH₂CH₂)₃Sn), 1.14-1.07 (m, 6 H, (CH₃CH₂CH₂CH₂)₃Sn), 0.88 (t,J=7.5 Hz, 9 H, (CH₃CH₂CH₂CH₂)₃Sn); HRMS (FAB), calcd. for C₁₆H₃₀FNSSn(M+H⁺) 408.1183, found 408.1169.

2-Ethoxy-4-ti-n-butylstannylthiazole 8d as illustrated in Scheme 4. Asolution of bromothiazole 21d (82 mg, 0.394 mmol, 1.0 equiv.) in ether(3.9 mL, 0.1 M) is treated with n-BuLi (289 μL, 1.5 M in hexanes, 0.433mmol, 1.1 equiv.) and tri-n-butyltin chloride (128 μL, 0.473 mmol, 1.2equiv.) according to the procedure described for the synthesis ofstannane 8s, to yield, after column chromatography (silica gel;pre-treated with Et₃N, hexanes), stannane 8d (161 mg, 98%). IR (film)ν_(max) 2956, 2927, 2870, 2851, 1504, 1472, 1258, 1257, 1232, 1211,1082, 1023, 960, 894, 872 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ6.65 (s, 1 H,ArH), 4.43 (q, J=7.0 Hz, 2 H, CH₃CH₂O), 1.61-1.53 (m, 6 H,(CH₃CH₂(CH₂)₂)₃Sn), 1.43 (t, J=7.0 Hz, 3 H, CH₃CH₂), 1.37-1.30 (m, 6 H,(CH₃CH₂CH₂CH₂)₃Sn), 1.08-1.04 (m, 6 H, (CH₃CH₂CH₂CH₂)₃Sn), 0.89 (t,J=7.5 Hz, 9 H, (CH₃CH₂CH₂CH₂)₃Sn); HRMS (FAB), calcd. for C₁₇H₃₃NOSSn(M+H⁺) 418.1380, found 418.1396.

2-Vinyl-4-tri-n-butylstannylthiazole 8q as illustrated in Scheme 5. Asolution of bromothiazole 21q (191 mg, 1.00 mmol, 1.0 equiv.) in ether(14.0 mL, 0.07 M), is treated with n-BuLi (804 μL, 1.5 M in hexanes,1.20 mmol, 1.2 equiv.) and tri-n-butyltin chloride (341 μL, 1.26 mmol,1.25 equiv.) according to the procedure described for the synthesis ofstannane 8s to yield, after column chromatography (silica gel;pre-treated with Et₃N, hexanes), stannane 8q (112 mg, 28%). R_(f)=0.63(silica gel, 17% ether in hexanes); IR (film) ν_(max) 2956, 2925, 2870,2850, 1459, 1377, 1205, 1080, 981, 913, 868 cm⁻¹; ¹H NMR (500 MHz,CDCl₃) δ7.21 (s, 1 H, ArH), 7.02 (dd, J=17.5, 11.0 Hz, 1 H, CH═CH₂),6.00 (d, J=17.5 Hz, 1 H, CHCH₂), 5.52 (d, J=11.0 Hz, 1 H, CH═CH₂),1.61-1.53 (m, 6 H, (CH₃CH₂(CH₂)₂)₃Sn), 1.37-1.27 (m, 6 H,(CH₃CH₂CH₂CH₂)₃Sn), 1.13-1.10 (m, 6 H, (CH₃CH₂CH₂CH₂)₃Sn), 0.88 (t,J=7.5 Hz, 9 H, (CH₃CH₂CH₂CH₂)₃Sn); HRMS (FAB), calcd. for C₁₇H₃₁NSSn(M+H⁺) 402.1279, found 402.1290.

2-Ethyl-4-tri-n-butylstannylthiazole 8r as illustrated in Scheme 5. Asolution of bromothiazole 21r (238 mg, 1.24 mmol, 1.0 equiv.) in ether(12.0 mL, 0.1M) at −78° C., is treated with n-BuLi (909 μL, 1.5 M inhexanes, 1.36 mmol, 1.1 equiv.) and tri-n-butyltin chloride (403 μL,1.49 mmol, 1.2 equiv.) according to the procedure described for thesynthesis of stannane 8s to yield, after column chromatography (silicagel; pre-treated with Et₃N, hexanes), stannane 8r (357 mg, 72%).R_(f)=0.64 (silica gel, CH₂Cl₂); IR (film) ν_(max) 2956, 2925, 2870,2852, 1464, 1376, 1292, 1174, 1072, 1033, 953, 875 cm⁻¹; ¹H NMR (400MHz, CDCl₃) δ7.18 (s, 1 H, ArH), 3.10 (q, J=7.6 Hz, 2 H, CH₃CH₂Ar),1.60-1.50 (m, 6 H, (CH₃CH₂(CH₂)₂)₃Sn), 1.39 (t, J=7.6 Hz, 3 H,CH₃CH₂Ar), 1.36-1.30 (m, 6 H, (CH₃CH₂CH₂CH₂)₃Sn), 1.13-1.08 (m, 6 H,(CH₃CH₂CH₂CH₂)₃Sn), 0.88 (t, J=7.3 Hz, 9 H, (CH₃CH₂CH₂CH₂)₃Sn); HRMS(FAB), calcd. for C₁₇H₃₃NSSn (M+H⁺) 404.1434, found 404.1416.

cis-Macrolactone 18h as illustrated in Scheme 3. A solution of vinyliodide 7 (10.0 mg, 0.020 mmol, 1.0 equiv.), stannane 8h (16.0 mg, 0.040mmol, 2.0 equiv.) and Pd(PPh₃)₄ (2.1 mg, 0.002 mmol, 0.1 equiv.) indegassed toluene (200 μL, 0.1 M) is heated at 100° C. for 20 min. Thereaction mixture is poured into saturated aqueous NaHCO₃—NaCl (5 mL) andextracted with EtOAc (2×5 mL). After drying the combined organicfractions (Na₂SO₄), evaporation of the solvents and purification bypreparative thin layer chromatography (500 mm silica gel plate, 50%EtOAc in hexanes), macrolactone 18h is obtained (7.5 mg, 76%).R_(f)=0.29 (silica gel, 50% EtOAc in hexanes); [α]²² _(D) −44.2 (c 0.60,CHCl₃); IR (thin film) ν_(max) 3387 (br), 2925, 2859, 1730, 1688, 1508,1461, 1256, 1183, 1150, 1061, 980, 755 cm⁻¹; ¹H NMR (500 MHz, CDCl₃)δ7.12 (s, 1 H, ArH), 6.61 (s, 1 H, CH═C(CH₃)), 5.45 (ddd, J=10.5, 10.5,4.5 Hz, 1 H, CH═CHCH₂), 5.38 (ddd, J=10.5, 10.5, 5.0 Hz, 1 H, CH═CHCH₂),5.31 (d, J=8.5 Hz, 1 H, CHOCO), 4.92 (d, J=4.0 Hz, 2 H, CH₂OH), 4.23(ddd, J=11.5, 5.5, 2.5 Hz, 1 H, (CH₃)₂CCH(OH)), 3.75-3.71 (m, 1 H,CHOH(CHCH₃)), 3.32 (d, J=5.5 Hz, 1 H, C(CH₃)₂CHOHO, 3.25 (t, J=4.0 Hz, 1H, CH₂OH), 3.13 (qd, J=7.0, 2.0 Hz, 1 H, CH₃CH(C═O)), 3.03 (d, J=2.0 Hz,1 H, CH₃CHCH(OH)CHCH₃), 2.68 (ddd, J=15.0, 9.5, 9.5 Hz, 1 H, ═CHCH₂CHO),2.50 (dd, J=15.0, 11.5 Hz, 1 H, CH₂COO), 2.35 (dd, J=15.0, 2.5 Hz, 1 H,CH₂COO), 2.31-2.24 (m, 1 H, ═CHCH₂CHO), 2.24-2.16 (m, 1 H), 2.09 (s, 3H, CH═CCH₃), 2.06-1.98 (m, 1 H), 1.82-1.73 (m, 1 H), 1.72-1.62 (m, 1 H),1.39-1.17 (m, 3 H), 1.33 (s, 3 H, C(CH₃)₂), 1.19 (d, J=7.0 Hz, 3 H,CH₃CH(C═O)), 1.08 (s, 3 H, C(CH₃)₂), 1.00 (d, J=7.0 Hz, 3 H, CH₃CHCH₂);HRMS (FAB), calcd. for C₂₆H₃₉NO₆S (M+Cs⁺) 626.1552, found 626.1530.

Epothilone E (3) as illustrated in Schemes 2 and 3. To a solution oflactone 18h (10.0 mg, 0.020 mmol, 1.0 equiv.) in methanol (600 μL, 0.03M) is added acetonitrile (32 μL, 0.606 mmol, 30 equiv.), KHCO₃ (10 mg,0.102 mmol, 5 equiv.) and hydrogen peroxide (27 μL, 35% w/w in water,0.303 mmol, 15 equiv.) and the reaction mixture stirred at 25° C. for 3h. Additional acetonitrile (32 μL, 0.606 mmol, 30 equiv.), KHCO₃ (10 mg,0.102 mmol, 5 equiv.) and hydrogen peroxide (27 μL, 35% w/w in water,0.303 mmol, 15 equiv.) are then added and stirring is continued for afurther 3 h. The reaction mixture is then passed directly through ashort plug of silica gel, eluting with ether, and the filtrate isconcentrated under reduced pressure. Preparative thin layerchromatography (250 mm silica gel plate, 50% EtOAc in hexanes) furnishesunreacted starting material 18h (5.0 mg, 50%) and epothilone E (3) (3.4mg, 33%). R_(f)=0.56 (silica gel, 66% EtOAc in hexanes); [α]²²_(D)=−27.5 (c 0.20, CHCl₃); IR (film) ν_(max) 3413, 2928, 2867, 1731,1689, 1462, 1375, 1257, 1152, 1061, 978, 756 cm⁻; ¹H NMR (600 MHz,CDCl₃) δ7.13 (s, 1 H, ArH), 6.61 (s, 1 H, CH═CCH₃), 5.46 (dd, J=8.1, 2.4Hz, 1 H, CHOCO), 4.94 (d, J=5.2 Hz, 2 H, CH₂OH), 4.16-4.12 (m, 1 H,(CH₃)₂CCH(OH)), 3.82-3.78 (m, 1 H, CHOH(CHCH₃)), 3.66 (bs, 1 H, OH),3.23 (qd, J=6.8, 5.2 Hz, 1 H, CH₃CH(C═O)), 3.04 (ddd, J=8.1, 4.5, 4.5Hz, 1 H, CH₂CH(O)CHCH₂), 2.91 (ddd, J=7.3, 4.5, 4.1 Hz, 1 H,CH₂CH(O)CHCH₂), 2.61 (t, J=5.2 Hz, 1 H, CH₂OH), 2.55 (dd, J=14.7, 10.4Hz, 1 H, CH₂COO), 2.48 (bs, 1 H, OH), 2.45 (dd, J=14.7, 3.2 Hz, 1 H,CH₂COO), 2.14-2.07 (m, 1 H, CH₂CH(O)CHCH₂), 2.11 (s, 3 H, CH═CCH₃), 1.91(ddd, J=15.1, 8.1, 8.1 Hz, 1 H, CH₂CH(O)CHCH₂), 1.78-1.66 (m, 2 H,CH₂CH(O)CHCH₂), 1.52-1.38 (m, 5 H), 1.36 (s, 3 H, C(CH₃)₂), 1.18 (d, 3H, J=6.8 Hz, CH₃CH(C═O)), 1.10 (s, 3 H, C(CH₃)₂), 1.01 (d, J=7.0 Hz, 3H, CH₃CHCH₂); HRMS (FAB), calcd. for C₂₆H₃₉NO₇S (M+H⁺) 510.2525, found510.2539.

cis-Macrolactone 18b as illustrated in Scheme 3. A solution of vinyliodide 7 (9.2 mg, 0.018 mmol, 1.0 equiv.), stannane 8b (10.7 mg, 0.036mmol, 2.0 equiv.) and Pd(PPh₃)₄ (2.1 mg, 0.0018 mmol, 0.1 equiv.) indegassed toluene (180 μL, 0.1 M) is heated at 100° C. for 40 min,according to the procedure described for the synthesis of lactone 18h,to yield, after preparative thin layer chromatography (250 mm silica gelplate, 75% ether in hexanes), macrolactone 18b (4.1 mg, 44%). R_(f)=0.50(silica gel, 50% EtOAc in hexanes); [α]²² _(D) −38.6 (c 0.21, CHCl₃); IR(thin film) ν_(max) 3444, 2925, 1732, 1682, 1259, 1037, 756 cm⁻¹; ¹H-NMR(500 MHz, CDCl₃) δ6.99 (s, 1 H, CH═C(CH₃)), 6.52 (bs, 1 H, ArH), 5.45(ddd, J=10.5, 10.5, 4.0 Hz, 2 H, CH═CHCH₂), 5.39 (ddd, J=10.5, 10.5, 4.0Hz, 1 H, CH═CHCH₂), 5.29 (d, J=8.0 Hz, 1 H, CHOCO), 4.20 (ddd, J=11.0,5.5, 2.5 Hz, 1 H, (CH₃)₂CCH(OH)), 3.75-3.73 (m, 1 H, CHOH(CHCH₃)), 3.13(qd, J=6.5, 2.0 Hz, 1 H, CH₃CH(C═O)), 2.98 (d, J=2.0 Hz, 1 H,CHOH(CHCH₃)), 2.93 (d, J=5.5 Hz, 1 H, (CH₃)₂CCH(OH)), 2.71 (ddd, J=15.0,10.0, 10.0 Hz, 1 H, CH═CHCH₂), 2.70 (s, 3 H, SCH₃), 2.51 (dd, J=15.5,11.5 Hz, 1 H, CH₂COO), 2.30 (dd, J=15.0, 2.5 Hz, 1 H, CH₂COO), 2.28-2.16(m, 2 H), 2.13 (d, J=1.0 Hz, 3 H, CH═CCH₃), 2.06-1.98 (m, 1 H),1.79-1.60 (m, 2 H), 1.40-1.06 (m, 3 H), 1.33 (s, 3 H, C(CH₃)₂), 1.19 (d,J=7.0 Hz, 3 H, CH₃CH(C═O)), 1.09 (s, 3 H, C(CH₃)₂), 1.00 (d, J=7.0 Hz, 3H, CH₃CHCH₂); HRMS (FAB), calcd. for C₂₆H₃₉NO₅S₂ (M+Cs⁺) 642.1324, found642.1345.

trans-Macrolactone 19b as illustrated in Scheme 3. A solution of vinyliodide 11 (6.9 mg, 0.014 mmol, 1.0 equiv.), stannane 8b (8.2 mg, 0.028mmol, 2.0 equiv.) and Pd(PPh₃)₄ (1.6 mg, 0.0014 mmol, 0.1 equiv.) indegassed toluene (140 μL, 0.1 M) is heated at 100° C. for 40 min,according to the procedure described for the synthesis of lactone 18h,to yield, after preparative thin layer chromatography (250 mm silica gelplate, 75% ether in hexanes), macrolactone 19b (5.0 mg, 72%). R_(f)=0.47(silica gel, 50% EtOAc in hexanes); [α]²² _(D) −32.9 (c 0.35, CHCl₃); IR(film) ν_(max) 3488, 2928, 1728, 1692, 1259, 1036, 800, 757 cm⁻¹; ¹H NMR(500 MHz, CDCl₃) δ7.00 (s, 1 H, ArH), 6.48 (s, 1 H, CH═CCH₃), 5.53 (ddd,J=15.0, 7.5, 7.5 Hz, 1 H, CH═CHCH₂), 5.40 (d, J=8.0 Hz, 1 H, CHOCO),5.39 (ddd, J=15.0, 7.5, 7.5 Hz, 1 H, CH═CHCH₂), 4.12 (ddd, J=11.0, 2.5,2.5 Hz, 1 H, (CH₃)₂CCHOH), 3.77-3.74 (m, 1 H, CHOH(CHCH₃)), 3.24 (m, 1H, CH═CHCH₂), 3.07 (m, 1 H, CH₃CH(C═O)), 2.70 (s, 3 H, SCH₃), 2.61 (d,J=3.5 Hz, 1 H, CHOH(CHCH₃)), 2.59-2.44 (m, 5 H), 2.19-2.12 (m, 1 H),2.13 (s, 3 H, CH═CCH₃), 2.02-1.94 (m, 1 H), 1.70-1.55 (m, 2 H),1.48-1.41 (m, 1 H), 1.29 (s, 3 H, C(CH₃)₂), 1.18 (d, J=7.0 Hz, 3 H,CH₃CH(C═O)), 1.08 (s, 3 H, C(CH₃)₂), 0.99 (d, J=7.0 Hz, 3 H, CH₃CHCH₂);HRMS (FAB), calcd. for C₂₆H₃₉NO₅S₂ (M+Cs⁺) 642.1324, found 642.1298.

cis-Macrolactone 18d as illustrated in Scheme 3. A solution of vinyliodide 7 (14 mg, 0.028 mmol, 1.0 equiv.), stannane 8d (14 mg, 0.055mmol, 2.0 equiv.) and PdCl₂(MeCN)₂ (2.0 mg, 0.008 mmol, 0.3 equiv.) indegassed DMF (280 μL, 0.1 M) is stirred at 25° C. for 20 h. Theresulting mixture is then concentrated under reduced pressure, filteredthrough silica, eluting with EtOAc, and purified by preparative thinlayer chromatography (250 mm silica gel plate, 50% ether in hexanes) tofurnish macrolactone 18d (12.5 mg, 89%). R_(f)=0.30 (silica gel, 66%ether in hexanes); [α]²² _(D) 70.2 (c 0.63, CHCl₃); IR (thin film)ν_(max) 3501 (br), 2934, 1732, 1688, 1526, 1472, 1386, 1232, 1150, 1091,1007 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ6.47 (s, 1 H, ArH), 6.33 (s, 1 H,CH═C(CH₃)), 5.43 (ddd, J=10.5, 10.5, 3.5 Hz, 1 H, CH═CHCH₂), 5.37 (ddd,J=10.5, 10.5, 4.5 Hz, 1 H, CH═CHCH₂), 5.26 (dd, J=9.5, 1.5 Hz, 1 H,CHOCO), 4.44 (q, J=7.0 Hz, 2 H, CH₃CH₂O), 4.18 (ddd, J=11.0, 5.5, 2.5Hz, 1 H, (CH₃)₂CCH(OH)), 3.73 (m, 1 H, CHOH(CHCH₃)), 3.12 (qd, J=7.0,2.0 Hz, 1 H, CH₃CH(C═O)), 2.98 (d, J=1.5 Hz, 1 H, OH), 2.95 (d, J=5.5Hz, 1 H, OH), 2.69 (ddd, J=15.0, 10.0, 10.0 Hz, 1 H, CH═CHCH₂CHO), 2.49(dd, J=15.5, 11.5 Hz, 1 H, CH₂COO), 2.36 (dd, J=15.5, 2.5 Hz, 1 H,CH₂COO), 2.23-2.16 (m, 3 H), 2.11 (s, 3 H, CH═C(CH₃)), 2.04-1.98 (m, 1H), 1.77-1.71 (m, 1 H), 1.70-1.61 (m, 1 H), 1.42 (t, J=7.0 Hz, 3 H,CH₃CH₂O), 1.38-1.16 (m, 2 H), 1.31 (s,3 H, C(CH₃)₂), 1.17 (d, J=7.0 Hz,3 H, CH₃CH(C═O)), 1.08 (s, 3 H, C(CH₃)₂), 0.99 (d, J=7.0 Hz, 3 H,CH₃CHCH₂); HRMS (FAB), calcd. for C₂₇H₄₁NO₆S (M+Cs⁺) 640.1709, found640.1732.

trans-Macrolactone 19d as illustrated in Scheme 3. A solution of vinyliodide 11 (14 mg, 0.028 mmol, 1.0 equiv.), stannane 8d (23 mg, 0.055mmol, 2.0 equiv.) and PdCl₂(MeCN)₂ (2.0 mg, 0.008 mmol, 0.3 equiv.) indegassed DMF (280 μL, 0.1 M) is stirred at 25° C. for 20 h, according tothe procedure described for the synthesis of macrolactone 18d to yield,after preparative thin layer chromatography (250 mm silica gel plate,50% EtOAc in hexanes), macrolactone 19d (12 mg, 86%). R_(f)=0.27 (silicagel, 66% ether in hexanes); [α]²² _(D) −28.0 (c 0.48, CHCl₃); IR (thinfilm) ν_(max) 3495 (br), 2930, 1732, 1690, 1526, 1472, 1233, 1017, 976cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ6.50 (s, 1 H, ArH), 6.30 (s, 1 H,CH═C(CH₃)), 5.57-5.51 (m, 1 H, CH═CHCH₂), 5.42-5.36 (m, 1 H, CH═CHCH₂),5.37 (dd, J=9.0, 2.5 Hz, 1 H, CHOCO), 4.46 (q, J=7.0 Hz, 2 H, CH₃CH₂O),4.10 (ddd, J=10.5, 3.5, 3.0 Hz, 1 H, (CH₃)₂CCH(OH)), 3.76-3.73 (m, 1 H,CHOH(CHCH₃)), 3.23 (qd, J=7.0, 4.5 Hz, 1 H, CH₃CH(C═O)), 3.07 (d, J=3.5Hz, 1 H, OH), 2.57-2.38 (m, 3 H), 2.56 (dd, J=15.5, 10.5 Hz, 1 H,CH₂COO), 2.47 (dd, J=15.5, 2.5 Hz, 1 H, CH₂COO), 2.18-2.16 (m, 1 H),2.13 (s, 3 H, CH═C(CH₃)), 2.03-1.94 (m, 1 H), 1.70-1.55 (m, 2 H),1.48-1.41 (m, 1 H), 1.44 (t, J=7.0 Hz, 3 H, CH₃CH₂O), 1.29 (s, 3 H,C(CH₃)₂), 1.27-1.16 (m, 1 H), 1.18 (d, J=7.0 Hz, 3 H, CH₃CH(C═O)), 1.08(s, 3 H, C(CH₃)₂), 0.98 (d, J=7.0 Hz, 3 H, CH₃CHCH₂); HRMS (FAB), calcd.for C₂₇H₄₁NO₆S (M+Cs⁺) 640.1709, found 640.1731.

tran-Macrolactone 19h as illustrated in Scheme 3. A solution of vinyliodide 11 (5.1 mg, 0.010 mmol, 1.0 equiv.), stannane 8h (8.0 mg, 0.020mmol, 2.0 equiv.) and Pd(PPh₃)₄ (1.1 mg, 0.001 mmol, 0.1 equiv.) indegassed toluene (100 μL, 0.1 M) is heated at 100° C. for 20 minaccording to the procedure described for the synthesis of macrolactone18h, to yield, after preparative thin layer chromatography (250 mmsilica gel plate, 50% EtOAc in hexanes), macrolactone 19h (4.3 mg, 88%).R_(f)=0.20 (silica gel, 50% EtOAc in hexanes); [α]²² _(D) −31.5 (c 0.60,CHCl₃); IR (thin film) ν_(max) 3410 (br), 2930, 1726, 1692, 1463, 1374,1255, 1180, 1064, 973 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ7.13 (s, 1 H, ArH),6.60 (s, 1 H, CH═C(CH₃)), 5.48 (ddd, J=15.0, 7.5, 7.5 Hz, 1 H,CH═CHCH₂), 5.40 (dd, J=5.5, 5.5 Hz, 1 H, CHOCO), 5.35 (ddd, J=15.0, 7.5,7.5 Hz, 1 H, CH═CHCH₂), 4.91 (d, J=7.0 Hz, 2 H, CH₂OH), 4.23 (ddd,J=9.5, 3.5, 3.0 Hz, 1 H, (CH₃)₂CCH(OH)), 3.74 (ddd, J=7.0, 5.0, 2.5 Hz,1 H, CHOH(CHCH₃)), 3.34 (t, J=7.0 Hz, 1 H, CH₂OH), 3.26 (qd, J=7.0, 7.0Hz, 1 H, CH₃CH(C═O)), 3.05 (d, J=3.5 Hz, 1 H, C(CH₃)₂CHOH), 3.00 (d,J=5.0 Hz, 1 H, CH₃CHCH(OH)CHCH₃), 2.56 (dd, J=15.5, 9.5 Hz, 1 H,CH₂COO), 2.47 (dd, J=15.5, 3.0 Hz, 1 H, CH₂COO), 2.58-2.45 (m, 1 H,═CHCH₂CH), 2.24-2.16 (m, 1 H, ═CHCH₂CH), 2.08 (s, 3 H, CH═CCH₃),1.98-1.90 (m, 1 H), 1.63-1.56 (m, 2 H), 1.54-1.46 (m, 1 H), 1.41-1.30(m, 1 H), 1.27 (s, 3 H, C(CH₃)₂), 1.20 (d, J=7.0 Hz, 3 H, CH₃CH(C═O)),1.07 (s, 3 H), C(CH₃)₂), 0.99 (d, J=7.0 Hz, 3 H, CH₃CHCH₂); HRMS (FAB),calcd. for C₂₆H₃₉NO₆S (M+Cs⁺) 626.1552, found 626.1536.

cis-Macrolactone 18j as illustrated in Scheme 3. A solution of vinyliodide 7 (12.5 mg, 0.025 mmol, 1.0 equiv.), stannane 8j (20 mg, 0.049mmol, 2.0 equiv.) and PdCl₂(MeCN)₂ (1.5 mg, 0.006 mmol, 0.2 equiv.) indegassed DMF (250 μL, 0.1 M) is stirred at 25° C. for 20 h, according tothe procedure described for the synthesis of macrolactone 18d, to yield,after preparative thin layer chromatography (250 mm silica gel plate,67% ether in hexanes) macrolactone 18j (9 mg, 74%). R_(f)=0.32 (silicagel, 50% EtOAc in hexanes); [α]²² _(D) −65.3 (c 0.45, CHCl₃); IR (thinfilm) ν_(max) 3406 (br), 2924, 2852, 1732, 1682, 1455, 1366, 1263, 1192,1148, 1096, 1043, 983, 881 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ7.21 (s, 1 H,ArH), 6.62 (s, 1 H, CH═C(CH₃)), 5.60 (d, J=47.0 Hz, 2 H, CH₂F), 5.45(ddd, J=10.5, 10.5, 4.0 Hz, 1 H, CH═CHCH₂), 5.38 (ddd, J=10.0, 10.0, 5.0Hz, 1 H, CH═CHCH₂), 5.31 (dd, J=10.0, 1.5 Hz, 1 H, CHOCO), 4.19 (ddd, 1H, J=11.0, 5.0, 2.5 Hz, 1 H, (CH₃)₂CCH(OH)), 3.73 (m, 1 H, CHOH(CHCH₃)),3.13 (qd, J=7.0, 2.0 Hz, 1 H, CH₃CH(C═O)), 2.97 (d, J=2.0 Hz, 1 H, OH),2.93 (d, J=5.5 Hz, 1 H, OH), 2.71 (ddd, J=15.0, 10.0, 10.0 Hz, 1 H,CH═CHCH₂CHO), 2.51 (dd, J=15.5, 11.5 Hz, 1 H, CH₂COO), 2.39 (dd, J=15.5,2.0 Hz, 1 H, CH₂COO), 2.29-2.22 (m, 1 H), 2.22-2.16 (m, 1 H), 2.11 (d,J=1.0 Hz, 3 H, CH═C(CH₃)), 2.06-1.99 (m, 1 H), 1.77-1.71 (m, 1 H),1.69-1.62 (m, 1 H), 1.38-1.16 (m, 3 H), 1.32 (s, 3 H, C(CH₃)₂), 1.18 (d,J=7.0 Hz, 3 H, CH₃CH(C═O)), 1.08 (s, 3 H, C(CH₃)₂), 1.00 (d, J=7.0 Hz, 3H, CH₃CHCH₂); HRMS (FAB), calcd. for C₂₆H₃₈FNO₅S (M+Cs⁺) 628.1509, found628.1530.

trans-Macrolactone 19j as illustrated in Scheme 3. A solution of vinyliodide 11 (15 mg, 0.030 mmol, 1.0 equiv.), stannane 8j (27 mg, 0.066mmol, 2.2 equiv.) and PdCl₂(MeCN)₂ (1.5 mg, 0.006 mmol, 0.2 equiv.) indegassed DMF (300 μL, 0.1 M) is stirred at 25° C. for 20 h, according tothe procedure described for the synthesis of macrolactone 18d, to yield,after preparative thin layer chromatography (250 mm silica gel plate,50% EtOAc in hexanes) macrolactone 19j (11 mg, 75%). R_(f)=0.17 (silicagel, 33% ether in hexanes); [α]²² _(D) −37.1 (c 0.55, CHCl₃); IR (thinfilm) ν_(max) 3508 (br), 2934, 1730, 1690, 1505, 1461, 1428, 1366, 1251,1196, 1150, 1041, 977 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ7.22 (s, 1 H, ArH),6.58 (s, 1 H, CH═C(CH₃)), 5.61 (d, J=47.0 Hz, 2 H, CH₂F), 5.55-5.50 (m,1 H, CH═CHCH₂), 5.41-5.35 (m, 2 H, CH═CHCH₂ and CHOCO), 4.15 (ddd,J=10.0, 3.5, 3.0 Hz, 1 H, (CH₃)₂CCH(OH)), 3.75-3.73 (m, 1 H,CHOH(CHCH₃)), 3.24 (qd, J=7.0, 4.5 Hz, 1 H, CH₃CH(C═O)), 3.05 (d, J=4.0Hz, 1 H, OH), 2.62 (d, J=4.0 Hz, 1 H, OH), 2.56 (dd, J=15.0, 10.5 Hz, 1H, CH₂COO), 2.49 (dd, J=15.5, 2.5 Hz, 1 H, CH₂COO), 2.49-2.44 (m, 2 H),2.20-2.13 (m, 1 H), 2.10 (s, 3 H, CH═C(CH₃)), 2.01-1.93 (m, 1 H),1.67-1.56 (m, 2 H), 1.49-1.43 (m, 1 H), 1.31-1.17 (m, 2 H), 1.28 (s, 3H, C(CH₃)₂), 1.18 (d, J=6.5 Hz, 3 H, CH₃CH(C═O)), 1.07 (s, 3 H,C(CH₃)₂), 0.98 (d, J=7.0 Hz, 3 H, CH₃CHCH₂); HRMS (FAB), calcd. forC₂₆H₃₈FNO₅S (M+Cs⁺) 628.1509, found 628.1487.

Silyl ether 25 as illustrated in Scheme 7. To a solution of alcohol 13(12.96 g, 54.4 mmol, 1.0 equiv.), in DMF (180 mL, 0.3 M) at 0° C., isadded imidazole (10.2 g, 150.0 mmol, 2.8 equiv.) followed bytert-butyldimethylchlorosilane (13.5 g, 89.8 mmol, 1.7 equiv.). Afterwarming to 25° C. over 7 h, the solvent is removed under reducedpressure and the resulting oil is partitioned between ether (200 mL) andsaturated aqueous NH₄Cl (200 mL). The aqueous layer is extracted withether (200 mL) and the combined organic extracts are washed with brine(550 mL), dried (MgSO₄) and concentrated under reduced pressure. Flashcolumn chromatography (silica gel, 0 to 5% EtOAc in hexanes) furnishessilyl ether 25 as an oil (16.03 g, 84%). R_(f)=0.48 (hexanes); [α]²²_(D) −17.5 (c 1.65, CHCl₃); IR (thin film) ν_(max) 2928, 2857, 1472,1361, 1278, 1252, 1082, 914, 836, 776, 677 cm⁻¹; ¹H NMR (500 MHz, CDCl₃)δ6.15 (s, 1 H, CH═CCH₃), 5.74-5.66 (m, 1 H, CH═CH₂), 5.03 (bm, 1 H,CH═CH₂), 5.01 (s, 1 H, CH═CH₂), 4.16 (dd, J=6.5, 6.5 Hz, 1 H, CHOH),2.25 (m, 1 H, CH₂═CHCH₂), 1.77 (s, 3 H, CH═CCH₃), 0.88 (s, 9 H,SiC(CH₃)₃), 0.04 (s, 3 H, Si(CH₃)₂), −0.01 (s, 3 H, Si(CH₃)₂);

Aldehyde 26 as illustrated in Scheme 7. To a solution of olefin 25 (16.0g, 45.3 mmol, 1.0 equiv.) in a mixture of THF (206 mL), t-BuOH (206 mL)and H₂O (41 mL) at 0° C. is added 4-methylmorpholine N-oxide (NMO) (5.84g, 49.8 mmol, 1.1 equiv.) followed by OsO₄ (5.2 mL, 2.5% w/v in t-BuOH,0.453 mmol, 0.01 equiv.). The mixture is vigorously stirred 13 h at 25°C. and then quenched with saturated aqueous Na₂SO₃ (125 mL). Theresulting solution is stirred for 2 h and then partitioned between EtOAc(150 mL) and water (150 mL). The organic phase is separated and theaqueous phase is extracted with EtOAc (2×200 mL). The combined organicextracts are dried (MgSO₄), filtered, and the solvents are removed underreduced pressure. Flash column chromatography (silica gel, 50 to 90%ether in hexanes) provides unreacted starting material (1.0 g, 6%) andthe desired diols as a ca. 1:1 mixture of diastereoisomers (15.5 g,89%). R_(f)=0.44 (silica gel, 50% EtOAc in hexanes); IR (thin film)ν_(max) 3387, 2952, 2928, 1252, 1080, 837, 777 cm⁻¹; ¹H NMR (500 MHz,CDCl₃) δ6.28 and 6.26 (singlets, 1 H total, CH═CCH₃), 4.47-4.42 (m, 1 H,CHOSi), 3.86-3.76 (m, 1 H, CHOH), 3.61-3.55 and 3.49-3.39 (m, 2 H total,CH₂OH), 3.33 and 3.15 (2 doublets, J=2.0 and 3.5 Hz, 1 H total, CHOH),2.46 and 2.45 (triplets, J=5.5 and 5.5 Hz, CH₂OH), 1.78 and 1.76(singlets, 3 H total), 1.63-1.60 and 1.58-1.53 (m, 2 H total, CH₂), 0.88and 0.87 (singlets, 9 H total, SiC(CH₃)₃), 0.08 and 0.07 (singlets, 3 Htotal, Si(CH₃)₂), 0.01 and 0.00 (singlets, 3 H total, Si(CH₃)₂); HRMS(FAB), calcd. for C₁₃H₂₇IO₃Si (M+Na⁺) 409.0672 found 409.0662.

The diols (obtained as described above) (23.3 g, 60.2 mmol, 1.0 equiv.)are dissolved in a mixture of MeOH (400 mL) and water (200 mL) and thesolution is cooled to 0° C. NaIO₄ (77.2 g, 361.1 mmol, 6.0 equiv.) isthen added portionwise over 5 min, and the resulting slurry is stirredvigorously for 30 min at 25° C. After completion of the reaction, themixture is partitioned between CH₂Cl₂ (500 mL) and water (500 mL) andthe organic phase is separated. The aqueous layer is extracted withCH₂Cl₂ (500 mL) and the combined organic extracts are washed with brine(1 L), dried (MgSO₄) and concentrated under reduced pressure. Flashcolumn chromatography (silica gel, 17 to 50% ether in hexanes) providesaldehyde 26 as an oil (19.6 g, 92%). R_(f)−0.35 (silica gel, 20% etherin hexanes); [α]²² _(D) −34.1 (c 2.8, CHCl₃); IR (thin film) ν_(max)2954, 2928, 2885, 2856, 1728, 1471, 1279, 1254, 1091, 838, 777, 677cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ9.73 (dd, J=2.5, 2.5 Hz, 1 H, CHO), 6.34(s, 1 H, CH═CCH₃), 4.70 (dd, J=8.0, 4.0 Hz, 1 H, CHOSi), 2.68 (ddd,J=16.0, 8.3, 2.5 Hz, 1 H, (CHO)CH₂), 2.44 (ddd, J=16.0, 4.0, 2.5 Hz, 1H, (CHO)CH₂), 1.80 (s, 3 H, CH═CCH₃), 0.85 (s, 9 H, SiC(CH₃)₃), 0.05 (s,3 H, Si(CH₃)₂), 0.01 (s, 3 H, Si(CH₃)₂); HRMS (FAB), calcd. forC₁₂H₂₃IO₂Si (M+Na⁺) 377.0410 found 377.0402.

Methyl ester 28 as illustrated in Scheme 7. A mixture of aldehyde 26(19.6 g, 55.2 mmol, 1.0 equiv.) and stabilized ylide 27 (50.2 g, 134.0mmol, 2.4 equiv.) [prepared from 4-bromo-1-butene by: (i) phosphoniumsalt formation; (ii) anion formation with KHMDS; and (iii) quenchingwith MeOC(O)Cl)] (see Marshall, J. A., et al., J. Org. Chem.51,1735-1741 (1986) and Bestmann, H. J., Angew. Chem. Int. Ed. Engl.1965, 645-60) in benzene (550 mL, 0.1 M) is heated at reflux for 1.5 h.After cooling to 25° C., the mixture is filtered and the solvent isremoved under reduced pressure. Flash column chromatography (silica gel,9 to 17% ether in hexanes) furnishes methyl ester 28 (24.5 g, 98%).R_(f)=0.37 (silica gel, 20% ether in hexanes); [α]²²−7.25 (c 1.6,CHCl₃); IR (thin film) ν_(max) 3078, 2952, 2920, 2856, 1720, 1462, 1434,1276, 1253, 1208, 1084, 836, 776, 672 cm⁻¹; ¹H NMR (600 MHz, CDCl₃)δ6.81 (dd, J=7.4, 7.4 Hz, 1 H, CH═CCOOCH₃), 6.22 (s, 1 H, CH═CCH₃),5.83-5.75 (m, 1 H, CH═CH₂), 4.99-4.98 (m, 1 H, CH═CH₂), 4.96 (m, 1 H,CH═CH₂), 4.22 (dd, J=7.5, 5.1 Hz, 1 H, CHOSi), 3.72 (s, 3 H, COOCH₃),3.05 (d, J=6.0 Hz, 2 H, CH₂C(CO₂Me)), 2.40 (ddd, J=15.0, 7.5, 7.5 Hz, 1H, CH₂CHOSi), 2.33 (ddd, J=15.0, 7.5, 5.1 Hz, 1 H, CH₂CHOSi), 1.77 (s, 3H, CH═CCH₃), 0.85 (s, 9 H, SiC(CH₃)₃), 0.02 (s, 3 H, Si(CH₃)₂), −0.02(s, 3 H, Si(CH₃)₂); HRMS (FAB), calcd. for C₁₈H₃₁IO₃Si (M+Cs⁺), 583.0142found 583.0159. Allylic alcohol 29 as illustrated in Scheme 7. Methylester 28 (24.5 g, 54.3 mmol, 1.0 equiv.) is dissolved in THF (280 mL)and the solution is cooled to −78° C. DIBAL (163.0 mL, 1 M in CH₂Cl₂,163.0 mmol, 3.0 equiv.) is added dropwise at −78° C. over 50 min, andthe reaction mixture is stirred for a further 80 min. The reactionmixture is quenched with saturated aqueous sodium-potassium tartrate(150 mL) and the resulting mixture is allowed to warm up to 25° C. over16 h. The organic layer is separated and the aqueous phase is extractedwith ether (3×250 mL). The combined organic extracts are washed withbrine (650 mL), dried (MgSO₄) and concentrated under reduced pressure.Flash column chromatography (silica gel, 17 to 50% ether in hexanes)furnishes alcohol 29 (22.9 g, 100%). R_(f)=0.11 (silica gel, 20% etherin hexanes); [α]²² _(D) −7.25 (c 1.6, CHCl₃); IR (thin film) ν_(max)3346, 3078, 2954, 2928, 2857, 1637, 1471, 1361, 1276, 1252, 1078, 1005,836, 775, 674, 558 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ6.16 (s, 1 H,CH═CCH₃), 5.81-5.73 (m, 1 H, CH═CH₂), 5.45 (dd, J=6.5, 6.5 Hz, 1 H,CH═CCH₂OH), 5.03 (m, 2 H, CH═CH₂), 4.16 (dd, J=6.5, 6.5 Hz, 1 H, CHOSi),4.02 (d, J=4.5 Hz, 2 H, CH₂OH), 2.85 (dd, J=15.0, 5.1 Hz, 1H,CH₂CH═CH₂), 2.84 (dd, J=15.0, 5.0 Hz, 1 H, CH₂CH═CH₂), 2.27 (ddd,J=15.0, 6.5, 6.5 Hz, 1 H, CH₂CHOSi), 2.25 (ddd, J=15.0, 6.5, 6.5 Hz, 1H, CH₂CHOSi), 1.78 (s, 3 H, CH═CCH₃), 0.88 (s, 9 H, SiC(CH₃)₃), 0.02 (s,3 H, Si(CH₃)₂), −0.02 (s, 3 H, Si(CH₃)₂); HRMS (FAB), calcd. forC17H₃₁IO₂Si (M+Cs⁺), 555.0192 found 555.0177.

Triphenylmethyl ether 30 as illustrated in Scheme 7. Alcohol 29 (23.5 g,55.7 mmol, 1.0 equiv.) is dissolved in DMF (300 mL, 0.15 M) and 4-DMAP(11.3 g, 92.5 mmol, 1.7 equiv.) and trityl chloride (22.1 g, 79.3 mmol,1.4 equiv.) are added. The reaction mixture is stirred at 80° C. for 21h, cooled to room temperature and the solvent is removed under reducedpressure. The resulting residue is purified by flash columnchromatography to afford the required ether 30 as an oil (35.3 g, 95%).R_(f)=0.88 (silica gel, 20% ether in hexanes); [α]²² _(D −)0.74 (c 0.3,CHCl₃); IR (thin film) ν_(max) 3058, 2927, 2854, 1488, 1470, 1448, 1250,1082, 836, 702, 632 cm⁻¹; ¹H NMR (600 MHz, CHCl₃) δ7.45-7.43 (m, 5 H,Ph), 7.32-7.21 (m, 10 H, Ph), 6.19 (s, 1 H, CH═CCH₃), 5.61 (m, 2 H,CH═CH₂, CH═CH₂), 4.87 (m, 2 H, CH═CH₂, CH(C)CH₂OTr), 4.19 (dd, J=6.8,6.8 Hz, 1 H, CHOSi), 3.46 (s, 2 H, CH₂OTr), 2.78 (dd, J=15.4, 6.7 Hz, 1H, CH₂CH═CH₂), 2.73 (dd, J=15.4, 6.3 Hz, 1 H, CH₂CH═CH₂), 2.33 (ddd,J=14.5, 6.8, 6.8 Hz, 1 H, CH₂CHOSi), 2.31 (ddd, J=14.5, 6.8, 6.8 Hz, 1H, CH₂CHOSi), 1.80 (s, 3 H, CH═CCH₃), 0.87 (s, 9 H, SiC(CH₃)₃), 0.04 (s,3 H, Si(CH₃)₂), 0.00 (s, 3 H, Si(CH₃)₂); HRMS (FAB), calcd. forC₃₆H₄₅IO₂Si (M+Cs⁺), 797.1288 found 797.1309.

Alcohol 31 as illustrated in Scheme 7. Olefin 30 (35.3 g, 53.1 mmol, 1.0equiv.) is dissolved in THF (53 mL, 1.0 M) and the solution is cooled to0° C. 9-BBN (149 mL, 0.5 M in THF, 74.5 mmol, 1.4 equiv.) is addeddropwise over 1.5 h, and the resulting mixture is stirred for 9 h at 0°C. Aqueous NaOH (106 mL of a 3 N solution, 319.0 mmol, 6.0 equiv.) isadded, followed by aqueous H₂O₂ (32 mL, 30% w/w in water, 319.0 mmol,6.0 equiv.). Stirring is continued for 1 h at 0° C., after which timethe reaction mixture is diluted with ether (500 mL) and water (500 mL).The organic layer is separated and the aqueous phase is extracted withether (2×500 mL). The combined organic extracts are washed with brine (1L), dried (MgSO₄) and concentrated under reduced pressure. Flash columnchromatography (silica gel, 9 to 50% ether in hexanes) furnishes primaryalcohol 31 (34.6 g, 95%). R_(f)=0.54 (silica gel, 60% ether in hexanes);[α]²² _(D) −3.5 (c 0.2, CHCl₃); IR (thin film) ν_(max) 3380, 3058, 3032,2926, 2855, 1489, 1449, 1278, 1251, 1078, 835, 706, 632 cm⁻¹; ¹H NMR(500 MHz, CDCl₃) δ7.47-7.45 (m, 5 H, Ph), 7.32-7.22 (m, 10 H, Ph), 6.22(s, 1 H, CH═CCH₃), 5.58 (dd, J=7.1, 7.1 Hz, 1 H, C═CHCH₂), 4.22 (dd,J=6.8, 6.0 Hz, 1 H, CHOSi), 3.52 (bm, 2 H, CH₂OH), 3.50 (s, 2 H,CH₂OTr), 2.33 (dd, J=14.5, 6.8, 6.8 Hz, 1 H, CH₂CHOSi), 2.28 (ddd,J=14.5, 6.8, 6.8 Hz, 1 H, CH₂CHOSi), 2.14 (m, 2 H, CH₂CH₂CH₂OH), 1.82(s, 3 H, CH═CCH₃), 1.46 (m, 2 H, CH₂CH₂OH), 0.90 (s, 9 H, SiC(CH₃)₃),0.06 (s, 3 H, Si(CH₃)₂), 0.02 (s, 3 H, Si(CH₃)₂); HRMS (FAB), calcd. forC₃₆H₄₇IO₃Si (M+Cs⁺), 815.1394 found 815.1430.

Iodide 32 as illustrated in Scheme 7. A solution of alcohol 31 (34.6 g,50.73 mmol, 1.0 equiv.) in a mixture of ether (380 mL) and MeCN (127 mL)is cooled to 0° C. Imidazole (17.3 g, 253.7 mmol, 5.0 equiv.) and PPh₃(33.3 g, 126.8 mmol, 2.5 equiv.) are then added and the mixture isstirred until all the solids have dissolved. Iodine (33.5 g, 131.9 mmol,2.6 equiv.) is added and the mixture is stirred for 45 min at 0° C. Thereaction is quenched by the addition of saturated aqueous Na₂S₂O₃ (150mL) and the layers are separated. The aqueous phase is then extractedwith ether (2×250 mL) and the combined organic extracts are washed withbrine (750 mL), dried (MgSO₄) and concentrated under reduced pressure.Flash column chromatography (silica gel, 5 to 9% ether in hexanes)furnishes iodide 32 (39.2 g, 97%). R_(f)=0.88 (silica gel, 60% ether inhexanes); [α]²² _(D) −2.9 (c 2.6, CHCl₃); IR (thin film) ν_(max) 3057,2926, 2855, 1481, 1448, 1251, 1083, 939, 836, 774, 706, 632 cm⁻¹; ¹H NMR(500 MHz, CHCl₃) δ7.49-7.45 (m, 5 H, Ph), 7.33-7.23 (m, 10 H, Ph), 6.23(s, 1 H, CH═CCH₃), 5.67 (dd, J=7.2, 7.1 Hz, 1 H, CH₂C═CH), 4.22 (dd,J=6.8, 6.8 Hz, 1 H, CHOSi), 3.51 (s, 2 H, CH₂OTr), 3.07 (dd, J=7.1, 7.0Hz, 2 H, CH₂I), 2.34 (ddd, J=14.5 6.8, 6.8 Hz, 1 H, CH₂CHOSi), 2.25(ddd, J=14.5, 6.8, 6.8 Hz, CH₂CHOSi), 2.13 (m, 2 H, CH₂CH₂CH₂l), 1.84(s, 3 H, CH═CCH₃), 1.75 (m, 2 H, CH₂CH₂CH₂I), 0.90 (s, 9 H, SiC(CH₃)₃),0.07 (s, 3 H, Si(CH₃)₂), 0.02 (s, 3 H, Si(CH₃)₂); HRMS (FAB), calcd. forC₃₆H₄₆I₂O₂Si (M+Cs⁺), 925.0411 found 925.0450.

Hydrazone 33 as illustrated in Scheme 7. Diisopropylamine (5.0 mL, 35.28mmol, 1.4 equiv.) is added to a solution of n-BuLi (22.0 mL, 1.6 M inhexanes, 35.28 mmol, 1.4 equiv.) in 32 mL of THF at 0° C. and stirredfor 1 h. The SAMP hydrazone of propionaldehyde (5.6 g, 32.76 mmol, 1.3equiv.) in THF (16 mL), is added to this freshly prepared solution ofLDA at 0° C. After stirring at that temperature for 16 h, the resultingyellow solution is cooled to −100° C., and a solution of iodide 32 (20.0g, 25.23 mmol, 1.0 equiv.) in THF (32 mL) is added dropwise over aperiod of 2 h. The mixture is allowed to warm to −20° C. over 20 h, andthen poured into saturated aqueous NH₄Cl (50 mL) and extracted withether (3×100 mL). The combined organic extract is dried (MgSO₄),filtered and evaporated. Purification by flash column chromatography onsilica gel (5 to 50% ether in hexanes) provides hydrazone 33 (15.0 g,71%) as a yellow oil. R_(f)=0.63 (silica gel, 40% ether in hexanes);[α]²² _(D) −22.7 (c 0.2, CHCl₃); IR (thin film) ν_(max) 3057, 2927,2854, 1489, 1448, 1251, 1078, 940, 836, 775, 706, 668, 632 cm⁻¹; ¹H NMR(500 MHz, CHCl₃) δ7.46-7.44 (m, 5 H, Ph), 7.31-7.21 (m, 10 H, Ph), 6.40(d, J=6.5 Hz, 1 H, N═CH), 6.21 (s, 1 H, CH═CCH₃), 5.50 (dd, J=7.0, 7,0Hz, 1 H, CH₂C═CH), 4.20 (dd, J=6.0, 6.0 Hz, 1 H, CHOSi), 3.54 (dd,J=9.2, 3.5 Hz, 1 H, CH₂OCH₃), 3.45 (s, 2 H, CH₂OTr), 3.41 (dd, J=9.5,7.0 Hz, 1 H, CH₂OCH₃), 3.37 (s, 3 H, CH₂OCH₃), 3.32-3.30 (m, 2 H, CH₂N),2.60-2.55 (m, 1 H), 2.34-2.20 (m, 3 H), 2.04-1.95 (m, 1 H), 1.98-1.73(m, 5 H), 1.82 (s, 3 H, CH═CCH₃), 1.38-1.21 (m, 4 H), 0.96 (d, J=6.9 Hz,3 H, CHCH₃), 0.89 (s, 9 H, SiC(CH₃)₃), 0.06 (S, 3 H, Si(CH₃)₂), 0.01 (s,3 H, Si(CH₃)₂; HRMS (FAB), calcd. for C₄₅H₆₃IN₂O₃Si (M+Cs⁺), 967.2707found 967.2740.

Nitrile 34 as illustrated in Scheme 7. Monoperoxyphthalic acid magnesiumsalt (MMPP.6H₂O, 80%, 52.4 g, 84.8 mmol, 2.5 equiv.) is addedportionwise over 10 min to a rapidly stirred solution of hydrazone 33(28.3 g, 33.9 mmol, 1.0 equiv.) in a mixture of MeOH (283 mL), THF (100mL) and pH 7 phosphate buffer (283 mL) at 0° C. The mixture is stirredat 0° C. for 1.5 h and then more THF (120 mL) is added in two portionsover 30 min to help dissolve the starting material. After stirring for afurther 1.5 h the reaction mixture is poured into saturated aqueoussolution of NaHCO₃ (750 mL) and the product is extracted with ether (750mL) and then EtOAc (2×750 mL). The combined organic extracts are washedwith brine (1 L), dried (MgSO₄) and concentrated under reduced pressure.Flash column chromatography (silica gel, 9 to 20% ether in hexanes)furnishes nitrile 34 as a colorless oil (21.8 g, 89%). R_(f)=0.44(silica gel, 20% ether in hexanes); [α]²² _(D) +2.9 (c 1.2, CHCl₃); IR(thin film) ν_(max) 3057, 2928, 2855, 2238, 1490, 1448, 1252, 1081, 836,775, 707, 632 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ7.47-7.45 (m, 5 H, Ph),7.33-7.23 (m, 10 H, Ph), 6.22 (s, 1 H, CH═CCH₃), 5.56 (dd, J=6.8, 6.8Hz, 1 H, CH₂C═CH), 4.21 (dd, J=6.8, 6.8 Hz, 1 H, CHOSi), 3.49 (s, 2 H,CH₂OTr), 2.48 (m, 1 H, CH(CH₃)), 2.29 (ddd, J=14.5, 6.8, 6.8 Hz, 1 H,CH₂CHOSi), 2.24 (ddd, J=14.5, 6.8, 6.8 Hz, 1 H, CH₂CHOSi), 2.07 (m, 2 H,CH₂(C)CH₂OTr)), 1.82 (s, 3 H, CH═CCH₃), 1.58-1.23 (m, 4 H), 1.24 (d,J=7.0 Hz, 3 H, CHCH₃), 0.90 (s, 9 H, SiC(CH₃)₃), 0.07 (s, 3 H,Si(CH₃)₂), 0.0 (s, 3 H, Si(CH₃)₂); HRMS (FAB), calcd. for C₃₉H₅₀INO₂Si(M+Cs⁺), 852.1710 found 852.1738.

Aldehyde 35 as illustrated in Scheme 7. Nitrile 34 (7.01 g, 9.74 mmol,1.0 equiv.) is dissolved in toluene (195 mL, 0.05 M) and cooled to −78°C. DIBAL (29.2 mL, 1.0 M in toluene, 29.2 mmol, 3.0 equiv.) is addeddropwise at −78° C. over 10 min. The reaction mixture is stirred at −78°C. until completion is verified by TLC (1 h). Methanol (10 mL) and HCl(10 mL, 1.0 N in water) are sequentially added and the resulting mixtureis brought up to 0° C. over 1 h. Ether (250 mL) and water (250 mL) areadded and the layers are separated. The aqueous phase is extracted withether (2×250 mL) and the combined organic extracts are washed with brine(500 mL), dried (MgSO₄) and concentrated under reduced pressure. Flashcolumn chromatography (silica gel, 17 to 33% ether in hexanes) affordsaldehyde 35 as an oil (6.18 g, 88%). R_(f)=0.51 (silica gel, 20% etherin hexanes); [α]²² _(D) +2.0 (c 0.3, CHCl₃); IR (thin film) ν_(max)3057, 2927, 2855, 1726, 1490, 1448, 1251, 1081, 836, 775, 707, 632 cm⁻¹;¹H NMR (500 MHz, CDCl₃) δ9.51 (d, J=1.9 Hz, 1 H, CHO), 7.46-7.45 (m, 5H, Ph), 7.32-7.22 (m, 10 H, Ph), 6.20 (s, 1 H, CH═CCH₃), 5.54 (dd,J=7.0, 7.0 Hz, 1 H, CH₂C═CH), 4.20 (dd, J=6.5, 6.0 Hz, 1 H, CHOSi), 3.47(s, 2 H, CH₂OTr), 2.34-2.20 (m, 3 H, CH₂CHOSi and CH(CH₃)), 2.04 (m, 2H, CH₂(C)CH₂OTr), 1.82 (s, 3 H, CH═CCH₃), 1.66 (m, 1 H), 1.30-1.19 (m, 3H), 1.02 (d, J=7.0 Hz, 3 H, CHCH₃), 0.89 (s, 9 H, SiC(CH₃)₃), 0.06 (s, 3H, Si(CH₃)₂), 0.00 (s, 3 H, Si(CH₃)₂); HRMS (FAB), calcd. forC₃₉H₅₁IO₃Si (M+Cs⁺), 855.1707 found 855.1672.

tris-(Silylethers) 37 and 38 as illustrated in Scheme 8. A solution ofketone 36 (see Nicolaou, K. C., et al., J. Am. Chem. Soc. 119, 7974-91(1997) (1.20 g, 2.99 mmol, 1.4 equiv.) in THF (4.3 mL) is added dropwiseover 5 min to a freshly prepared solution of LDA [diisopropylamine (424μL, 3.03 mmol, 1.45 equiv.) is added to n-BuLi (2.00 mL, 1.52 M inhexanes, 3.04 mmol, 1.45 equiv.) at 0° C., and after 5 min THF (4.3 mL)is added] at −78° C. After stirring for 1.5 h at −78° C., the solutionis allowed to warm up to 40° C. over a period of 30 min. The reactionmixture is then cooled to −78° C., and a solution of aldehyde 35 (1.51g, 2.09 mmol, 1.0 equiv.) in THF (12.5 mL) is added dropwise over 15min. The resulting mixture is stirred for 1 h at −78° C., and thenquenched by dropwise addition of saturated aqueous AcOH (3.1 mL of a 1 Msolution in THF, 3.10 mmol, 1.5 equiv.). The mixture is then warmed to25° C. and partitioned between ether (25 mL) and saturated aqueous NH₄Cl(25 mL). The aqueous phase is extracted with ether (3×25 mL) and thecombined organic extracts are dried (MgSO₄) and concentrated underreduced pressure. Flash column chromatography (silica gel, 4 to 20%ether in hexanes) provides unreacted ketone (502 mg, 42%), undesiredaldol product 38 (705 mg, 27%) and a mixture of desired aldol product 37and unreacted aldehyde 35 [1.136 g, (ca. 9:1 ratio of 37:35 by ¹H NMR)](i.e. 39% yield of 37). This mixture is used directly in the next step.37: (major) (obtained as a colorless oil from a mixture containing 35,by flash column chromatography silica gel, (10 to 17% EtOAc in hexanes).R_(f)=0.22 (silica gel, 10% ether in hexanes); [α]²² _(D) −20.0 (c 0.3,CHCl₃); IR (thin film) ν_(max) 3486, 2954, 2928, 2856, 1682, 1472, 1448,1253, 1090, 994, 836, 775, 706, 668, 632 cm⁻¹; ¹H NMR (600 MHz, CDCl₃)δ7.45-7.43 (m, 5 H, Ph), 7.30-7.19 (m, 10 H, Ph) 6.19 (sm 1 H, CH═CCH₃),5.51 (dd, J=7.0, 6.9 Hz, 1 H, C═CHCH₂), 4.18 (dd, J=6.3, 6.2 Hz, 1 H,CHOSi), 3.88 (dd, J=7.5, 2.6 Hz, 1 H, CHOSi), 3.65 (m, 1 H, CH₂OSi),3.59 (m, 1 H, CH₂OSi), 3.46 (d, J=11.2 Hz, 1 H, CH₂OTr), 3.43 (d, J=11.2Hz, 1 H, CH₂OTr), 3.27 (m, 1 H, CH₃CH(C═O)), 3.22 (d, J=9.3 Hz, 1 H,CHOH), 2.32-2.18 (m, 2 H, C═CHCH₂CHOSi) 2.00 (m, 2 H, CH₂(C)CH₂OTr),1.80 (s, 3 H, CH═C(CH₃)), 1.66 (m, 2 H), 1.46 (m, 2 H), 1.27 (m, 1 H,CH(CH₃), 1.19 (s, 3 H, C(CH₃)₂), 1.07 (s, 3 H, C(CH₃)₂), 0.99 (d, J=6.8Hz, 3 H, CH(CH₃)), 0.89 (s, 9 H, SiC(CH₃)₃), 0.87 (s, 9 H, SiC(CH₃)₃),0.86 (s, 9 H, SiC(CH₃)₃), 0.71 (d, J=6.7 Hz, 3 H, CH(CH₃)), 0.10 (s, 3H, Si(CH₃)₂), 0.07 (s, 3 H, Si(CH₃)₂), 0.04 (s, 3 H, Si(CH₃)₂), 0.03 (s,6 H, Si(CH₃)₂), −0.01 (s, 3 H, Si(CH₃)₂); HRMS (FAB), calcd. forC₆₀H₉₇IO₆Si₃ (M+Cs⁺), 1257.4692 found 1257.4639. 38: (minor) Colorlessoil; R_(f)=0.38 (silica gel, 20% ether in hexanes); [α]²² _(D) −11.9 (c2.9, CHCl₃); IR (thin film) ν_(max) 3501, 2954, 2930, 2856, 1682, 1469,1254, 1088, 836, 776, 705, 670 cm⁻¹; ¹H NMR (500 MHz, CHCl₃) δ7.46-7.44(m, 5 H, Ph), 7.31-7.21 (m, 10 H, Ph), 6.21 (s, 1 H, CH═CCH₃), 5.52 (dd,J=7.0, 6.9Hz, 1 H, C═CHCH₂), 4.20 (dd, J=6.5, 6.5 Hz, 1 H, CHOSi), 3.88(dd, J=7.5, 2.5 Hz, 1 H, CHOSi), 3.67 (m, 1 H, CH₂OSi), 3.60 (m, 1 H,CH₂OSi), 3.46 (s, 2 H, CH₂OTr), 3.30-3.21 (m, 2 H, CHOH, CH₃CH(C═O)),2.30-2.25 (m, 2 H, C═CHCH₂CHOSi), 2.05-1.93 (m, 2 H, CH₂C(CH₂OTr)═CH),1.81 (s, 3 H, CH═C(CH₃)), 1.63 (m, 1 H, CH(CH₃), 1.45 (m, 2 H), 1.24 (m,2 H), (s, 3 H, C(CH₃)₂), 1.05 (s, 3 H, C(CH₃)₂), 1.01 (d, J=6.9 Hz, 3 H,CH(CH₃)), 0.92 (s, 18 H, SiC(CH₃)₃), 0.89 (s, 9 H, SiC(CH₃)₃), 0.88(obscured d, 3 H, CH(CH₃)), 0.88 (s, 18 H, SiC(CH₃)₃), 0.11 (s, 3 H,Si(CH₃)₂), 0.07 (s, 3 H, Si(CH₃)₂), 0.06 (s, 3 H, Si(CH₃)₂), 0.04 (s, 6H, Si(CH₃)₂), 0.01 (s, 3 H, Si(CH₃)₂); HRMS (FAB), calcd. forC₆₀H₉₇IO₆Si₃ (M+Cs⁺), 1257.4692 found 1257.4749.

tetra-(Silylether) 39 as illustrated in Scheme 8. Alcohol 37 (1.136 g ofa 9:1 mixture with aldehyde 35, 0.933 mmol, 1.0 equiv.) is dissolved inCH₂Cl₂ (5.0 mL), cooled to −20° C. and treated with 2,6-lutidine (470μL, 4.04 mmol, 4.3 equiv.) and tert-butyidimethylsilyltrifluoromethanesulfonate (695 μL, 3.03 mmol, 3.2 equiv.). The mixtureis then stirred for 2.5 h with slow warming to 0° C. The reaction isthen quenched with saturated aqueous NaHCO₃ (25 mL) and the aqueousphase is extracted with ether (3×25 mL). The combined organic extractsare washed with brine (250 mL), dried (MgSO₄) and concentrated underreduced pressure. Flash column chromatography (silica gel, 4 to 9% etherin hexanes) furnishes tetra-(silylether) 39 as a colorless oil (1.04 g,90%). R_(f)=0.91 (silica gel, 20% ether in hexanes); [α]²² _(D) −16.8 (c0.7, CHCl₃); IR (thin film) ν_(max) 3058, 2951, 2856, 1693, 1471, 1253,1079, 1004, 836, 706 cm⁻¹; ¹H NMR (600 MHz, CDCl₃) δ7.46-7.43 (m, 5 H,Ph), 7.29-7.19 (m, 10 H, Ph), 6.19 (s, 1 H, CH═CCH₃), 5.49 (dd, J=7.0,7.0 Hz, 1 H, C═CHCH₂), 418 (dd, J=6.3, 6.1 Hz, 1 H, CHOSi), 3.85 (dd,J=7.6, 2.5 Hz, 1 H, CHOSi), 3.70 (dd, J=6.7, 2.0 Hz, 1 H, CHOSi), 3.67(ddd, J=9.6, 4.8, 4.8 Hz, 1 H, CH₂OSi), 3.59 (ddd, J=9.7, 7.9. 7.9 Hz, 1H, CH₂OSi), 3.45 (d, J=11.2 Hz, 1 H, CH₂OTr), 3.42 (d, J=11.2 Hz, 1 H,CH₂OTr), 3.08 (qd, J=6.8, 6.8 Hz, 1 H, CH₃CH(C═O)), 2.27 (ddd, J=14.4,7.2, 7.2 Hz, 1 H, C═CHCH₂CHOSi), 2.23 (ddd, J=14.5, 6.2, 6.2 Hz, 1 H,C═CHCH₂CHOSi), 1.97 (m, 2 H, CH₂C(CH₂OTr)═CH), 1.79 (s, 3 H, CH═C(CH₃)),1.57 (m, 1 H), 1.46 (m, 1 H), 1.25 (m, 3 H), 1.17 (s, 3 H, C(CH₃)₂),1.01 (d, J=6.8 Hz, 3 H, CH(CH₃)), 0.95 (s, 3 H, C(CH₃)₂), 0.87 (s, 18 H,SiC(CH₃)₃), 0.86 (s, 18 H, SiC(CH₃)₃), 0.09-−0.03 (m, 24 H, Si(CH₃)₂);HRMS (FAB), calcd. for C₆₆H₁₁₁IO₆Si₄ (M+Cs⁺), 1371.5557 found 1371.5523.

Alcohol 40 as illustrated in Scheme 8. To a solution of tetra-silylether 39 (180 mg, 0.145 mmol) in THF (1.5 mL) at 0° C. is added HFepyr.in pyridine/THF mixture (prepared from a stock solution containing 420μL HF.pyridine, 1.14 mL pyridine and 2.00 mL THF) (1.5 mL) and theresulting solution is stirred for 2 h at 0° C. More HF.pyr. inpyridine/THF mixture (0.5 mL) is then added and stirring is continuedfor additional 1 h at 0° C. The reaction is quenched by careful additionof saturated aqueous NaHCO₃ and the product is extracted with EtOAc(3×25 mL). The combined organic extracts are then dried (MgSO₄) andconcentrated under reduced pressure. Flash chromatography (silica gel30% ether in hexanes) fumishes alcohol 40 as a pale yellow oil (137 mg,84%). R_(f)=0.36 (silica gel, 40% ether in hexanes); [α]²² _(D) −26.0 (c0.3, CHCl₃); IR (thin film) ν_(max) 3422, 2928, 2855, 1690, 1490. 1471,1448, 1360, 1252, 1086, 1004, 986, 836, 774, 706 cm⁻¹; ¹H NMR (600 MHz,CDCl₃) δ7.44-7.42 (m, 5 H, Ph), 7.29-7.20 (m, 10 H, Ph), 6.19 (s, 1 H,CH═CCH₃), 5.49 (dd, J=7.1, 7.1 Hz, 1 H, C═CHCH₂), 4.17 (dd, J=6.2, 6.0Hz, 1 H, CHOSi), 4.03 (dd, J=6.6, 3.7 Hz, 1 H, CHOSi), 3.73 (dd, J=7.2,1.7 Hz, 1 H, CHOSi), 3.65 m, 2 H, CH₂OH), 3.45 (d, J=11.7 Hz, 1 H,CH₂OTr), 3.42 (d, J=11.7 Hz, 1 H, CH₂OTr), 3.06 (qd, J=6.9, 6.9 Hz, 1 H,CH₃CH(C═O)), 2.28 (ddd, J=14.7, 7.3, 7.3 Hz, 1 H, C═CHCH₂CHOSi), 2.22(ddd, J=14.7, 6.3, 6.3 Hz, 1 H, C═CHCH₂CHOSi), 1.98 (m, 2 H,CH₂C(CH₂OTr)═CH), 1.79 (s, 3 H, CH═C(CH₃)), 1.56 (m, 2 H), 1.24 (m, 3H), 1.18 (s, 3 H, C(CH₃)₂), 1.03 (d, J=6.9 Hz, 3 H, CH(CH₃)), 0.97 (s, 3H, C(CH₃)₂), 0.87 (3 singlets, 27 H, SiC(CH₃)₃), 0.81 (d, J=6.7 Hz, 3 H,CH(CH₃)), 0.10 (s, 3 H, Si(CH₃)₂), 0.04 (s, 9 H, Si(CH₃)₂), 0.03 (s, 3H, Si(CH₃)₂), 0.00 (s, 3 H, Si(CH₃)₂); HRMS (FAB), calcd. forC₆₀H₉₇IO₆Si₃ (M+Cs⁺), 1257.4692 found 1257.4780.

Aldehyde 41 as illustrated in Scheme 8. To a solution of oxalyl chloride(150 μL, 1.72 mmol, 2.0 equiv.) in CH₂Cl₂ (10 mL) at −78° C. is addeddropwise DMSO (247 μL, 3.48 mmol, 4.0 equiv.). After stirring for 10 minat −78° C., a solution of alcohol 40 (960 mg, 0.853 mmol, 1.0 equiv.) inCH₂Cl₂ (10 mL) is added dropwise. The resulting solution is stirred at−78° C. for 1 h, and then Et₃N (714 μL, 5.12 mmol, 6.0 equiv.) is addedand the reaction mixture is allowed to warm up to 25° C. over 30 min.Water (30 mL) is added, and the product is extracted with ether (3×40mL). The combined organic extracts are dried (MgSO₄) and thenconcentrated under reduced pressure. Flash column chromatography (silicagel, 17 to 50% ether in hexanes) furnishes aldehyde 41 as a colorlessoil (943 mg, 98%). R_(f)=0.74 (silica gel, 40% ether in hexanes); [α]²²_(D) −10.8 (c 0.1, CHCl₃); IR (thin film) ν_(max) 2928, 2855, 1728,1690, 1471, 1448, 1260, 1252, 1085, 987, 836, 774, 706 cm⁻¹; ¹H NMR (600MHz, CDCl₃) δ9.74 (dd, J=2.4, 1.5 Hz, 1 H, CHO), 7.44-7.42 (m, 5 H, Ph),7.29-7.20 (m, 10 H, Ph), 6.19 (s, 1 H, CH═CCH₃), 5.49 (dd, J=7.0, 6.8Hz, 1 H, C═CHCH₂), 4.44 (dd, J=6.3, 5.0 Hz, 1 H, CHOSi), 4.18 (dd,J=6.9, 6.4 Hz, 1 H, CHOSi), 3.70 (dd, J=7.2, 1.8 Hz, 1 H, CHOSi), 3.45(d, J=11.4 Hz, 1 H, CH₂OTr), 3.42 (d, J=11.4 Hz, 1 H, CH₂OTr), 3.05 (qd,J=7.0, 7.0 Hz, 1 H, CH₃CH(C═O)), 2.49 (ddd, J=17.0, 4.5, 1.4 Hz,CH₂CHO), 2.38 (ddd, J=17.0, 5.4, 2.8 Hz, 1 H, CH₂CHO), 2.27 (ddd,J=14.0, 7.1, 7.1 Hz, 1 H, C═CHCH₂CHOSi), 2.23 (ddd, J=14.5, 6.5, 6.5 Hz,1 H, C═CHCH₂CHOSi), 1.98 (m, 2 H, CH₂C(CH₂OTr)═CH), 1.79 (s, 3 H,CH═C(CH₃)), 1.27 (m, 4 H), 1.19 (s, 3 H, C(CH₃)₂), 1.12 (m, 1 H), 1.00(d, J=6.8 Hz, 3 H, CH(CH₃)), 0.98 (s, 3 H, C(CH₃)₂), 0.87 (s, 27 H,Si(CH₃)₃), 0.80 (d, J=6.7 Hz, 3 H, CH(CH₃)), 0.07 (s, 3 H, Si(CH₃)₂),0.04 (s, 3 H, Si(CH₃)₂), 0.03 (s, 3 H, Si(CH₃)₂), 0.03 (s, 3 H,Si(CH₃)₂), 0.02 (s, 3 H, Si(CH₃)₂), 0.00 (s, 3 H, Si(CH₃)₂); HRMS (FAB),calcd. for C₆₀H₉₅IO₆Si₃ (M+Cs⁺), 1255.4536 found 1255.4561.

Carboxylic acid 42 as illustrated in Scheme 8. To a solution of aldehyde41 (943 mg, 0.839 mmol, 1.0 equiv.) in t-BuOH (38.5 mL) and H₂O (8.4 mL)is added 2-methyl-2-butene (31.5 mL, 2 M in THF, 63.0 mmol, 75 equiv.)and NaH₂PO₄ (250 mg, 2.08 mmol, 2.5 equiv.) followed by NaClO₂ (380 mg,4.20 mmol, 5.0 equiv.) and the resulting mixture is stirred at 25° C.for 40 min. The volatiles are then removed under reduced pressure andthe residue is partitioned between EtOAc (40 mL) and brine (40 mL) andthe layers are separated. The aqueous phase is then extracted with EtOAc(3×40 mL), and the combined organic extracts are dried (MgSO₄) and thenconcentrated under reduced pressure. Flash column chromatography (silicagel, 60% ether in hexanes) furnishes carboxylic acid 42 as an oil (956mg, 100%). R_(f)=0.47 (silica gel, 40% ether in hexanes); [α]²² _(D)−19.6 (c 0.2, CHCl₃); IR (thin film) ν_(max) 3389, 2930, 2856, 1711,1469, 1254, 1085, 988, 835, 775, 705 cm⁻¹; ¹H NMR (600 MHz, CDCl₃)δ7.44-7.43 (m, 5 H, Ph), 7.29-7.20 (m, 10 H, Ph), 6.19 (s, 1 H,CH═CCH₃), 5.49 (dd, J=7.3, 7.1 Hz, 1 H, C═CHCH₂), 4.34 (dd, J=6.4, 3.3Hz, 1 H, CHOSi), 4.18 (dd, J=6.2, 6.2 Hz, 1 H, CHOSi), 3.72 (dd, J=7.2,1.7 Hz, 1 H, CHOSi), 3.45 (d, J=11.4 Hz, 1 H, CH₂OTr), 3.41 (d, J=11.4Hz, 1 H, CH₂OTr), 3.07 (qd, J=7.0, 7.0 Hz, 1 H, CH₃CH(C═O)), 2.46 (dd,J=16.3, 3.1 Hz, 1 H, CH₂CO₂H), 2.32-2.18 (m, 3 H, CH₂CO₂H andC═CHCH₂CHOSi), 1.97 (m, 2 H, CH₂C(CH₂OTr)═CH), 1.80 (s, 3 H, CH═C(CH₃)),1.31-1.19 (m, 5 H), 1.19 (s, 3 H, C(CH₃)₂), 1.02 (d, J=6.9 Hz, 3 H,CH(CH₃)), 0.99 (s, 3 H, C(CH₃)₂), 0.87 (s, 27 H, Si(CH₃)₃), 0.80 (d,J=6.8 Hz, 3 H, CH(CH₃)), 0.07, (s, 3 H, Si(CH₃)₂), 0.04 (s, 3 H,Si(CH₃)₂), 0.04 (s, 3 H, Si(CH₃)₂), 0.03 (s, 3 H, Si(CH₃)₂), 0.03 (s, 3H, Si(CH₃)₂), 0.02 (s, 3 H, Si(CH₃)₂), 0.00 (s, 3 H, Si(CH₃)₂); HRMS(FAB), calcd. for C₆₀H₉₅IO₇Si₃ (M+Cs⁺), 1271.4485 found 1271.4550.

Hydroxy acid 43 as illustrated in Scheme 8. A solution of carboxylicacid 42 (956 mg, 0.839 mmol, 1.0 equiv.) in THF (17 mL) at 0° C. istreated with TBAF (5.0 mL, 1.0 M in THF, 5.00 mmol, 6.0 equiv.) and themixture is allowed to warm to 25° C. over 19 h. The reaction is thenquenched by the addition of saturated aqueous NH₄Cl (40 mL) and theproduct is extracted with EtOAc (3×40 mL). The combined organic extractsare dried (MgSO₄) and concentrated under reduced pressure. Flash columnchromatography (silica gel, 5% MeOH in CH₂Cl₂) furnishes hydroxy acid 43as a yellow oil (817 mg, 95%). R_(f)=0.27 (silica gel, 5% MeOH inCH₂Cl₂); [α]²² _(D) −1.4 (c 0.2, CHCl₃); IR (thin film) ν_(max) 3364,3057, 2938, 2856, 1712, 1694, 1469, 1254, 1086, 1053, 988, 836, 776,734, 705 cm⁻¹; ¹H NMR (600 MHz, CDCl₃) δ7.43-7.42 (m, 5 H, Ph),7.30-7.21 (m, 10 H, Ph), 6.32 (s, 1 H, CH═CCH₃), 5.46 (dd, J=7.2, 7.2Hz, 1 H, C═CHCH₂), 4.35 (dd, J=6.3, 3.2 Hz, 1 H, CHOH), 4.21 (dd, J=6.4,6.3 Hz, 1 H, CHOSi), 3.73 (dd, J=7.3, 1.2 Hz, 1 H, CHOSi), 3.52 (d,J=12.1 Hz, 1 H, CH₂OTr), 3.48 (d, J=12.1 Hz, 1 H, CH₂OTr), 3.06 (m, 2 H,CH₃CH(C═O) and OH), 2.45 (dd, J=16.4, 3.0 Hz, 1 H, CH₂CO₂H), 2.35 (m, 2H, C═CHCH₂CHOH), 2.29 (dd, J=16.4, 6.5 Hz, 1 H, CH₂CO₂H), 2.07-1.94 (m,2 H, CH₂C(CH₂OTr)═CH), 1.85 (s, 3 H, CH═C(CH₃)), 1.71 (m, 1 H), 1.39 (m,1 H, CH(CH₃)), 1.27 (m, 3 H), 1.18 (s, 3 H, C(CH₃)₂), 1.02 (obscured d,3 H, CH(CH₃)), 1.02 (s, 3 H, C(CH₃)₂), 0.87 (s, 18 H, Si(CH₃)₃), 0.81(d, J=6.8 Hz, 3 H, CH(CH₃)), 0.09 (s, 3 H, Si(CH₃)₂), 0.07 (s, 3 H,Si(CH₃)₂), 0.04 (s, 3 H, Si(CH₃)₂), 0.02 (s, 3 H, Si(CH₃)₂); HRMS (FAB),calcd. for C₅₄H₈₁IO₇Si₂ (M+Cs⁺), 1157.3620 found 1157.3669.

Macrolactone 44 as illustrated in Scheme 8: To a solution of hydroxyacid 43 (1.06 g, 1.04 mmol, 1.0 equiv.) in THF (15 mL, 0.07 M) is addedEt₃N (870 μL 0.24 mmol, 6.0 equiv.) and 2,4,6-trichlorobenzoyl chloride(390 μL, 2.50 mmol, 2.4 equiv.). The reaction mixture is stirred at 0°C. for 1.5 h, and then added slowly over a period of 2 h via syringepump to a solution of 4-DMAP (280 mg, 2.29 mmol, 2.2 equiv.) in toluene(208 mL, 0.005 M based on 43) at 75° C. The mixture is stirred at thattemperature for an additional 0.5 h and is then concentrated underreduced pressure. The resulting residue is filtered through a plug ofsilica gel eluting with 50% ether in hexanes. Flash columnchromatography (silica gel, 17% ether in hexanes) furnishes macrolactone44 as a colorless foam (877 mg, 84%). R_(f)=0.19 (10% ether in hexanes);[α]²² _(D) −7.4 (c 0.2, CHCl₃); IR (thin film) ν_(max) 2929, 2855, 1742,1695, 1468, 1381, 1253, 1156, 1065, 985, 834, 774, 733, 706 cm⁻¹; ¹H NMR(600 MHz, CDCl₃) δ7.44-7.42 (m, 5 H, Ph), 7.29-7.20 (m, 10 H, Ph), 6.39(s, 1 H, CH═CCH₃), 5.51 (dd, J=9.5, 6.8 Hz, 1 H, C═CHCH₂), 5.07 (d,J=9.3 Hz, 1 H, CHOCO), 4.02 (d, J=9.2 Hz, 1 H, CHOSi), 3.82 (d, J=8.9Hz, 1 H, CHOSi), 3.46 (d, J=11.5 Hz, 1 H, CH₂OTr), 3.42 (d, J=11.5 Hz, 1H, CH₂OTr), 2.95 (dq, J=8.7, 7.0 Hz, 1 H, CH₃CH(C═O)), 2.72 (m, 2 H,C═CHCH₂CHO and CH₂COO), 2.54 (dd, J=16.2, 9.7 Hz, 1 H, CH₂COO), 2.29 (m,1 H, C═CHCH₂CHO), 2.12 (dd, J=14.3, 5.1 Hz, 1 H, CH₂C(CH₂OTr)═CH), 1.98(m, CH₂C(CH₂OTr)═CH), 1.88 (s, 3 H, CH═C(CH₃)), 1.44-1.23 (m, 5 H), 1.18(s, 3 H, C(CH₃)₂), 1.10 (s, 3 H, C(CH₃)₂), 1.07 (d, J=6.8 Hz, 3 H,CH(CH₃)), 0.92 ((s, 9 H, Si(CH₃)₃), 0.82 (d, J=6.9 Hz, 3 H, CH(CH₃)),0.72 (s, 9 H, Si(CH₃)₃), 0.08 (s, 3 H, Si(CH₃)₂), 0.05 (s, 3 H,Si(CH₃)₂), 0.05 (s, 3 H, Si(CH₃)₂), −0.32 (s, 3 H, Si(CH₃)₂); HRMS(FAB), cald. for C₅₄H₇₉IO₆Si₂ (M+Cs⁺), 1139.3514 found 1139.3459.

Triol 24 as illustrated in Scheme 8. To a solution of macrolactone 44(608 mg, 0.604 mmol, 1.0 equiv.) in THF (45 mL) at 0° C. is addedHF.pyr. (15 mL). The resulting mixture is allowed to warm up to 25° C.over 15 h and is then cooled to 0° C. and quenched by careful additionof saturated aqueous NaHCO₃ (50 mL). The product is then extracted withEtOAc (3×50 mL), and the combined organic extracts are dried (MgSO₄) andthen concentrated under reduced pressure. Flash column chromatography(silica gel, 60% EtOAc in hexanes) furnishes triol 24 as a colorlessfoam (280 mg, 86%). R_(f)=0.32 (silica gel, 60% EtOAc in hexanes); [α]²²_(D) −32.1 (c 0.2, CHCl₃); IR (thin film) ν_(max) 3413, 2923, 2857,1731, 1686, 1461, 1379, 1259, 1148, 1046, 737 cm⁻¹; ¹H NMR (600 MHz,CDCl₃) δ6.43 (s, 1 H, CH═CCH₃), 5.38 (dd, J=9.7, 5.4 Hz, 1 H, C═CHCH₂),5.29 (dd, J=8.8, 1.9 Hz, 1 H, CHOCO), 4.08 (m, 1 H, CHOH), 4.06 (d,J=13.0 Hz, 1 H, CH₂OH), 4.00 (d, J=13.0 Hz, 1 H, CH₂OH), 3.69 (dd,J=3.5. 3.4Hz, 1 H, CHOH), 3.12 (qd, J=6.9, 3.1 Hz, 1 H, CH₃CH(C═O)),2.76 (bs, 1 H, OH), 2.67 (ddd, J=15.0, 9.7, 9.7 Hz, 1 H, C═CHCH₂CHO),2.45 (dd, J=15.4, 10.6 Hz, 1 H, CH₂COO), 2.38 (bs, 1 H, OH), 2.33 (dd,J=15.4, 3.0 Hz, 1 H, CH₂COO), 2.21 (m, 2 H, CH₂C(CH₂OH)═CH), 2.06 (m, 1H, C═CHCH₂CHO), 1.87 (s, 3 H, CH═C(CH₃)), 1.71 (m, 1 H), 1.66 (m, 1 H),1.32 (s, 3 H, C(CH₃)₂), 1.29-1.24 (m, 3 H), 1.17 (d, J=6.9 Hz, 3 H,CH(CH₃)), 1.08 (s, 3 H, C(CH₃)₂), 0.99 (d, J=7.0 Hz, 3 H, CH(CH₃)); HRMS(FAB), calcd. for C₂₃H₃₇IO₆ (M+Cs⁺), 669.0689 found 669.0711.

Macrolactone 45 as illustrated in Scheme 9. A solution of vinyl iodide24 (55 mg, 0.103 mmol, 1.0 equiv.), stannane 8j (84 mg, 0.207 mmol, 2.0equiv.) and PdCl₂(MeCN)₂ (4 mg, 0.015 mmol, 0.15 equiv.) in degassed DMF(1 mL, 0.1 M) is stirred at 25° C. for 33 h, according to the proceduredescribed for the synthesis of macrolactone 18d, to yield, afterpreparative thin layer chromatography (250 mm silica gel plates, 80%EtOAc in hexanes), starting vinyl iodide 24 (21 mg, 39%) andmacrolactone 45 (30 mg, 56%). R_(f)=0.48 (silica gel, 80% EtOAc inhexanes); [α]²² _(D) −48.3 (c 0.2, CHCl₃); IR (thin fiim) ν_(max) 3372,2924, 2860, 1731, 1682, 1454, 1384, 1252, 1148, 1040, 979, 735 cm⁻¹; ¹HNMR (600 MHz, CHCl₃) δ7.21 (s, 1 H, ArH), 6.61 (s, 1 H, C═CCH₃), 5.58(d, J=47.0 Hz, 2 H, CH₂F), 5.45 (dd, J=9.8, 5.3 Hz, 1 H, C═CHCH₂), 5.26(dd, J=9.4, 2.0 Hz, 1 H, CHOCO), 4.23 (dd, J=10.9, 2.4 Hz, 1 H, CHOH),4.08 (d, J=13.1 Hz, 1 H, CH₂OH), 4.01 (d, J=13.1 Hz, 1 H, CH₂OH), 3.70(dd, J=4.2, 2.7 Hz, 1 H, CHOH), 3.16 (qd, J=6.8, 2.6 Hz, 1 H,CH₃CH(C═O)), 2.94 (bs, 1 H, OH), 2.69 (ddd, J=15.2, 9.6, 9.6 Hz, 1 H,C═CHCH₂CHO), 2.46 (dd, J=14.8, 10.9 Hz, 1 H, CH₂COO), 2.36-2.24 (m, 2 H,CH₂C(CH₂OH)═CH), 2.30 (dd, J=14.8, 2.6 Hz, 1 H, CH₂COO), 2.09 (s, 3 H,CH═C(CH₃)), 2.07 (m, 1 H, C═CHCH₂CHO), 1.77-1.58 (m, 5 H), 1.33 (s, 3 H,C(CH₃)₂), 1.17 (d, J=6.9 Hz, 3 H, CH(CH₃)), 1.06 (s, 3 H, C(CH₃)₂), 1.00(d, J=7.0 Hz, 3 H, CH(CH₃)); HRMS (FAB), calcd. for C₂₂H₄₀FNO₆S (M+Cs⁺),658.1615 found 658.1644.

Macrolactone 46 as illustrated in Scheme 9. A solution of vinyl iodide24 (32 mg, 0.060 mmol, 1.0 equiv.), stannane 8p (28 mg, 0.101 mmol, 1.7equiv.) and PdCl₂(MeCN)₂ (1.7 mg, 0.07 mmol, 0.1 equiv.) in degassed DMF(650 μL, 0.1 M) is stirred at 25° C. for 20 h, according to theprocedure described for the synthesis of macrolactone 18d, to yield,after preparative thin layer chromatography (250 mm silica gel plates,80% EtOAc in hexanes), starting vinyl iodide 24 (6 mg, 19%) andmacrolactone 46 (17 mg, 54%). R_(f)=0.37 (silica gel, 80% EtOAc inhexanes); [α]²² _(D) −48.7 (c 0.15, CHCl₃); IR (thin film) ν_(max) 3402,2931, 2874, 1731, 1686, 1533, 1458, 1420, 1383, 1242, 1150, 1048, 1007,979 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ6.50 (s, 1 H, ArH), 6.36 (s, 1 H,CH+CCH₃), 5.45 (dd, J=10.0, 5.0 Hz, 1 H, C═CHCH₂), 5.23 (dd, J=9.5, 1.5Hz, 1 H, CHOCO), 4.24 (bd, J=11.0 Hz, 1 H, CHOH), 4.11-3.68 (m, 1 H,CH₂OH), 4.07 (s, 3 H, OCH₃), 4.01 (d, J=13.0 Hz, 1 H, CH₂OH), 3.71 (dd,J=4.0, 2.5 Hz, 1 H, CHOH), 3.30 (bs, 1 H, OH), 3.16 (qd, J=7.0, 2.5 Hz,1 H, CH₃CH(C═O)), 3.00 (bs, 1 H, OH), 2.68 (ddd, J=15.0, 10.0, 9.5 Hz, 1H, C═CHCH₂CHO), 2.46 (dd, J=15.0, 11.0 Hz, 1 H, CH₂COO), 2.30-2.20 (m, 2H, CH₂C(CH₂OH)═CH), 2.29 (dd, J=15.0, 3.0 Hz, 1 H, CH₂COO), 2.11-2.04(m, 1 H, C═CHCH₂CHO), 2.11 (s, 3 H, CH═C(CH₃)), 1.83-1.61 (m, 4 H),1.41-1.25 (m, 1 H), 1.33 (s, 3 H, C(CH₃)₂), 1.18 (d, J=7.0 Hz, 3 H,CH(CH₃)), 1.07 (s, 3 H, C(CH₃)₂), 1.01 (d, J=7.0 Hz, 3 H, CH(CH₃)); HRMS(FAB), calcd. for C₂₇H₄₁NO₇S (M+Cs⁺), 656.1658 found 656.1675.

Macrolactone 47 as illustrated in Scheme 9. A solution of vinyl iodide24 (41 mg, 0.076 mmol, 1.0 equiv.), stannane 8r (61 mg, 0.151 mmol, 2.0equiv.) and PdCl₂(MeCN)₂ (4 mg, 0.015 mmol, 0.2 equiv.) in degassed DMF(760 μL, 0.1 M) is stirred at 25° C. for 21 h, according to theprocedure described for the synthesis of macrolactone 18d, to yield,after preparative thin layer chromatography (250 mm silica gel plates,80% EtOAc in hexanes), starting vinyl iodide 24 (6 mg, 15%) andmacrolactone 47 (20.5 mg, 51%). R_(f)=0.41 (silica gel, 80% hEtOAc inhexanes); [α]²² _(D) −86.0 (c 0.25, CHCl₃); IR (thin film) ν_(max) 3387,2968, 2936, 2874, 1733, 1685, 1458, 1381, 1253, 1149, 1050, 1003, 912cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ6.97 (s, 1 H, ArH), 6.63 (s, 1 H,CHCCH₃), 5.43 (dd, J=9.0, 5.5 Hz, 1 H, C═CHCH₂), 5.25 (dd, J=8.5, 2.0Hz, 1 H, CHOCO), 4.32 (ddd, J 11.0, 5.5, 2.5 Hz, 1 H, CHOH), 4.12-4.07(m, 2 H, CH₂OH and OH), 4.02 (d, J=11.0 Hz, 1 H, CH₂₀H), 3.69 (dd,J=2.0, 2.0 Hz, 1 H, CHOH), 3.16 (qd, J=7.0, 2.5 Hz, 1 H, CH₃CH(C═O)),3.08 (bs, 1 H, OH), 2.98 (q, J=7.0 Hz, 2 H, CH₂CH₃), 2.61 (ddd, J=15.0,9.0, 9.0 Hz, 1 H, C═CHCH₂CHO), 2.46 (dd, J=14.5, 11.0 Hz, 1 H, CH₂COO),2.38 (dd, J=15.0, 4.0 Hz, 1 H, CH₂C(CH₂OH)═CH), 2.31-2.25 (m, 1 H,CH₂C(CH₂OH)═CH), 2.23 (dd, J=14.5, 2.5 Hz, 1 H, CH₂COO), 2.17-2.07 (m, 1H, C═CHCH₂CHO), 2.04 (s, 3 H, CH═C(CH₃)), 1.97 (bs, 1 H, OH), 1.78-1.61(m, 3 H), 1.38-1.23 (m, 2 H), 1.37 (q, J=7.0 Hz, 3 H, CH₂CH₃), 1.35 (s,3 H, C(CH₃)₂), 1.18 (d, J=7.0 Hz 3 H, CH(CH₃)), 1.05 (s, 3 H, C(CH₃)₂),1.01 (d, J=7.0 Hz, 3 H, CH(CH₃)); HRMS (FAB), calcd. for C₂₈H₄₃NO₆S(M+Na⁺), 544.2709 found 544.2724.

Macrolactone 48 as illustrated in Scheme 9. A solution of vinyl iodide24 (26 mg, 0.048 mmol, 1.0 equiv.), stannane 8h (29 mg, 0.072 mmol, 1.5equiv.) and PdCl₂(MeCN)₂ (1.5 mg, 0.006 mmol, 0.1 equiv.) in degassedDMF (480 μL, 0.1 M) is stirred at 25° C. for 15 h, according to theprocedure described for the synthesis of macrolactone 18d, to yield,after preparative thin layer chromatography (250 mm silica gel plates,EtOAc), starting vinyl iodide 24 (10.5 mg, 40%) and macrolactone 48(10.5 mg, 41%). R_(f)=0.27 (silica gel, EtOAc); [α]²² _(D) 31 43.0 (c0.14, CHCl₃); IR (thin film) ν_(max) 3388, 2924, 2851, 1732, 1682, 1462,1384, 1251, 1185, 1150, 1067 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ7.13 (s, 1H, ArH), 6.63 (s, 1 H, CH═CCH₃), 5.45 (dd, J=9.0, 6.0 Hz, 1 H, C═CHCH₂),5.27 (bd, J=7.0 Hz, 1 H, CHOCO), 4.29 (dd, J=11.0, 2.5 Hz, 1 H, CHOH),4.09 (d, J=13.0 Hz, 1 H, CH₂OH), 4.00 (d, J=13.0 Hz, 1 H, CH₂OH), 3.68(dd, J=4.0, 2.5 Hz, 1 H, CHOH), 3.15 (qd, J=6.5, 2.5 Hz, 1 H,CH₃CH(C═O)), 2.99 (bs, 1 H, OH), 2.65 (ddd, J=15.0, 9.0, 9.0 Hz, 1 H,C═CHCH₂CHO), 2.46 (dd, J=14.5, 11.0 Hz, 1 H, CH₂COO), 2.39-2.33 (m, 1 H,CH₂C(CH₂OH)═CH), 2.26 (dd, J=14.5, 2.5 Hz, 1 H, CH₂COO), 2.26-2.20 (m, 1H, CH₂C(CH₂OH)═CH), 2.14-2.10 (m, 1 H, C═CHCH₂CHO), 2.07 (s, 3 H,CH═C(CH₃)), 1.99-1.61 (m, 4 H), 1.42-1.24 (m, 2 H), 1.33 (s, 3 H,C(CH₃)₂), 1.16 (d, J=7.0 Hz, 3 H, CH(CH₃)), 1.04 (s, 3 H, C(CH₃)₂), 1.00(d, J=7.0 Hz, 3 H, CH(CH₃)); HRMS (FAB), calcd. for C₂₇H₄₁NO₇S (M+Cs⁺),656.1658 found 656.1677.

Macrolactone 49 as illustrated in Scheme 9. A solution of vinyl iodide24 (37 mg, 0.069 mmol, 1.0 equiv.), stannane 8q (47 mg, 0.117 mmol, 1.7equiv.) and Pd(PPh₃)₄ (10 mg, 0.009 mmol, 0.13 equiv.) in degassedtoluene (780 μL, 0.1 M) is heated at 100° C. for 2 h according to theprocedure described for the synthesis of macrolactone 18h, to yield,after preparative thin layer chromatography (250 mm silica gel plates,80% EtOAc in hexanes), macrolactone 49 (5.5 mg, 15%). R_(f)=0.35 (silicagel, 80% EtOAc in hexanes); [α]²² _(D) −48.1 (c 0.27, CHCl₃); IR (thinfilm) ν_(max) 3403, 2930, 2873, 1732, 1686, 1462, 1381, 1291, 1266,1250, 1149, 1004, 980, 937 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ7.04 (s, 1 H,ArH), 6.85 (dd, J=17.5, 11.0 Hz, 1 H, CH═CH₂), 6.61 (s, 1 H, CH═CCH₃),6.05 (d, J=17.5 Hz, 1 H, CH═CH₂), 5.56 (d, J=11.0 Hz, 1 H, CH═CH₂), 5.45(dd, J=10.0, 5.5 Hz, 1 H, C═CHCH₂), 5.26 (dd, J=9.5, 2.0 Hz, 1 H,CHOCO), 4.29 (ddd, J=11.0, 6.0, 2.5 Hz, 1 H, CHOH), 4.09 (dd, J=13.0,6.5 Hz, 1 H, CH₂OH), 4.02 (dd, J=13.0, 6.0 Hz, 1 H, CH₂OH), 3.71 (ddd,J=4.5, 2.5, 2.5 Hz, 1 H, CHOH), 3.54 (d, J=6.0 Hz, 1 H, OH), 3.17 (qd,J=7.5, 2.0 Hz, 1 H, CH₃CH(C═O)), 3.02 (d, J=2.0 Hz, 1 H, OH), 2.68 (ddd,J=15.0, 10.0, 9.0 Hz, 1 H, C═CHCH₂CHO), 2.45 (dd, J=14.5, 11.0 Hz, 1 H,3 H, CH₂COO), 2.37-2.31 (m, 1 H, CH₂C(CH₂OH)═CH), 2.30-2.24 (m, 1 H,CH₂C(CH₂OH)═CH), 2.28 (dd, J=15.0, 3.5 Hz, 1 H, CH₂COO), 2.14-2.07 (m, 1H, C═CHCH₂CHO), 2.09 (d, J=1.0 Hz, 1 H, CH═C(CH₃)), 1.79-1.60 (m, 4 H),1.39-1.25 (m, 2 H), 1.35 (s, 3 H, C(CH₃)₂), 1.18 (d, J=7.0 Hz, 3 H,CH(CH₃)), 1.07 (s, 3 H, C(CH₃)₂), 1.02 (d, J=7.0 Hz, 3 H, CH(CH₃)); HRMS(FAB), calcd. for C₂₈H₄₁NO₆S (M+Cs⁺), 652.1709 found 652.1693.

Fluoride 50 as illustrated in Scheme 9: A solution of triol 45 (3.6 mg,0.007 mmol, 1.0 equiv.) in CH₂Cl₂ (0.1 mL, 0.07 M) at −78° C. is treatedwith DAST (11 μL of a 0.7 M solution in CH₂Cl₂, 0.008 mmol, 1.1 equiv.)and the mixture is stirred at −78° C. for 10 min. The reaction is thenquenched by the addition of saturated aqueous NaHCO₃ (500 μL) and themixture is allowed to warm to 25° C. The product is then partitionedbetween saturated aqueous NaHCO₃ (5 mL) and CH₂Cl₂ (5 mL) and the layersare separated. The aqueous phase is extracted with CH₂Cl₂ (2×5 mL) andthe combined organic extracts are dried (MgSO₄) and then concentratedunder reduced pressure. Preparative thin layer chromatography (250 mmsilica gel plate, 40% EtOAc in hexanes) furnishes fluoride 50 (2.1 mg,58%). R_(f)=0.39 (silica gel, 50% EtOAc in hexanes); [α]²² _(D) −34.4 (c0.09, CHCl₃); IR (thin film) ν_(max) 3413, 2919, 2849, 1725, 1684, 1465,1381, 1290, 1250, 1150, 1041, 979, 872 cm⁻¹; ¹H NMR (600 MHz, CDCl₃)δ7.22 (s, 1 H, ArH), 6.62 (s, 1 H, CH═CCH₃), 5.60 (d, J=47.0 Hz, 2 H,ArCH₂F), 5.56-5.52 (m, 1 H, C═CHCH₂), 5.27 (dd, J=9.5, 2.0 Hz, 1 H,CHOCO), 4.79 (dd, J=82.2, 10.8 Hz, 1 H, CH═CCH₂F), 4.71 (dd, J=81.8,10.8 Hz, 1 H, CH═CCH₂F), 4.24 (dd, J=10.9, 2.6 Hz, 1 H, CHOH), 3.70 (dd,J=4.3, 2.5 Hz, 1 H, CHOH), 3.15 (qd, J=6.8, 2.5 Hz, 1 H, CH₃CH(C═O)),3.00-2.85 (m, 1 H, OH), 2.71 (m, 1 H, C═CHCH₂CHO), 2.46 (dd, J=14.9,11.0 Hz, 1 H, CH₂COO), 2.38-2.29 (m, 2 H, CH₂C(CH₂OH)═CH), 2.30 (dd,J=14.9, 2.8 Hz, 1 H, CH₂COO), 2.15-2.09 (m, 1 H, C═CHCH₂CHO), 2.11 (d,J=1.0 Hz, CH═C(CH₃)), 1.80-1.50 (m, 4 H), 1.37-1.29 (m, 2 H), 1.33 (s, 3H, C(CH₃)₂), 1.18 d, J=6.8 Hz, 3 H, CH(CH₃)), 1.06 (s, 3 H, C(CH₃)₂),1.01 (d, J=7.1 Hz, 3 H, CH(CH₃)); HRMS (FAB calcd. for C₂₇H₃₉F₂NO₅S(M+H⁺), 528.2595 found 528.2610.

Fluoride 51 as illustrated in Scheme 9. A solution of triol 46 (8.2 mg,0.016 mmol, 1.0 equiv.) in CH₂Cl₂ (200 μL, 0.08 M) at −78° C. is treatedwith DAST (0.025 mL, 0.019 mmol, 1.2 equiv.) and the resulting mixtureis stirred at −78° C. for 10 min according to the procedure describedfor the synthesis of fluoride 50, to yield, after preparative thin layerchromatography (250 mm silica gel plates, 30% EtOAc in hexanes),fluoride 51 (3.5 mg, 43%). R_(f)=0.57 (silica gel, 60% EtOAc inhexanes); [α]²² _(D) −4.7 (c 0.11, CHCl₃); IR (thin film) ν_(max) 3418,2925, 2852, 1734, 1686, 1535, 1461, 1415, 1383, 1334, 1241, 1150, 1045,976 cm⁻¹; ¹H NMR (400 MHz, CHCl₃) δ6.51 (s, 1 H, ArH), 6.37 (s, 1 H,CH═CCH₃), 5.55-5.51 (m, 1 H, C═CHCH₂), 5.22 (dd, J=10.0, 2.0 Hz, 1 H,CHOCO), 4.81 (dd, J=74.0, 11.0 Hz, 1 H, CH═CCH₂F), 4.71 (dd, J=73.0,11.0 Hz, 1 H, CH═CCH₂F), 4.26 (dd, J=11.0, 2.5 Hz, 1 H, CHOH), 4.09 (s,3 H, CH₃O), 3.71 (dd, J=4.5, 2.0 Hz, 1 H, CHOH), 3.17 (qd, J=7.0, 2.5Hz, 1 H, CH₃CH(C═O)), 3.01-2.95 (m, 1 H, OH), 2.76-2.68 (m, 1 H,C═CHCH₂CHO), 2.47 (dd, J=14.5, 11.0 Hz, 1 H, CH₂COO), 2.37-2.27 (m, 2 H,CH₂C(CH₂OH)═CH), 2.29 (dd, J=14.5, 2.5 Hz, 1 H, CH₂COO), 2.17-2.11 (m, 1H, C═CHCH₂CHO), 2.14 (s, 3 H, CH═C(CH₃)), 1.80-1.50 (m, 4 H), 1.40-1.22(m, 2 H), 1.34 (s, 3 H, C(CH₃)₂), 1.19 (d, J=7.0 Hz, 3 H, CH(CH₃)), 1.08(s, 3 H, C(CH₃)₂), 1.03 (d, J=7.0 Hz, 3 H, CH(CH₃)); HRMS (FAB), calcd.for C₂₇H₄₀FNO₆S (M+H⁺), 526.2639 found 526.2625.

Fluoride 52 as illustrated in Scheme 9. A solution of triol 47 (12.5 mg,0.024 mmol, 1.0 equiv.) in CH₂Cl₂ (500 μL, 0.05 M) at −78° C. is treatedwith DAST (250 RL, 0.1 M in CH₂Cl₂, 0.025 mmol, 1.05 equiv.) and theresulting mixture is stirred at −78° C. for 10 min according to theprocedure described for the synthesis of fluoride 50, to yield, afterpreparative thin layer chromatography (250 mm silica gel plates, 60%EtOAc in hexanes), fluoride 52 (5.1 mg, 41%). R_(f)=0.19 (silica gel,50% EtOAc in hexanes); [α]²² _(D) −68.6 (c 0.22, CHCl₃); IR (thin film)ν_(max) 3504, 2969, 2935, 2877, 1736, 1687, 1461, 1369, 1290, 1250,1148, 1068, 1044, 1008, 976 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ6.98 (s, 1 H,ArH), 6.60 (s, 1 H, CH═CCH₃), 5.56-5.52 (m, 1 H, C═CHCH₂), 5.23 (dd,J=10.0, 2.0 Hz, 1 H, CHOCO), 4.80 (dd, J=73.0, 10.5 Hz, 1 H, CH═CCH₂F),4.71 (dd, J=72.5, 10.5 Hz, 1 H, CH═CCH₂F), 4.33 (ddd, J=11.0, 5.5, 2.5Hz, 1 H, CHOH), 3.71 (ddd, J=5.0, 2.5, 2.0 Hz, 1 H, CHOH), 3.71 (d,J=6.0 Hz, 1 H, CHOH), 3.17 (qd, J=7.0, 2.0 Hz, 1 H, CH₃CH(C═O)), 3.07(m, 1 H, OH), 4.51 (q, J=7.5 Hz, 2 H, CH₂CH₃), 2.70 (ddd, J=15.0, 10.0,2.0 Hz, 1 H, C═CHCH₂CHO), 2.45 (dd, J=14.5, 11.0 Hz, 1 H, CH₂COO),2.39-2.28 (m, 2 H, CH₂C(CH₂OH)═CH), 2.26 (dd, J=14.5, 2.5 Hz, 1 H,CH₂COO), 2.17-2.10 (m, 1 H, C═CHCH₂CHO), 2.08 (d, J=1.5 Hz, 3 H,CH═C(CH₃)), 1.80-1.67 (m, 3 H), 1.39 (t, J=7.5 Hz, 3 H, CH₂CH₃),1.39-1.24 (m, 2 H), 1.35 (s, 3 H, C(CH₃)₂), 1.19 (d, J=7.0 Hz, 3 H,CH(CH₃)), 1.07 (s, 3 H, C(CH₃)₂), 1.03 (d, J=7.0 Hz, 3 H, CH(CH₃)); HRMS(FAB), calcd. for C₂₈H₄₂FNO₅S (M+Cs⁺), 656.1822 found 656.1843.

Fluoride 53 as illustrated in Scheme 9. A solution of triol 49 (6.0 mg,0.0151 mmol, 1.0 equiv.) in CH₂Cl₂ (1.5 mL, 0.01 M) at −78° C. istreated with DAST (0.20 mL, 0.08 M in CH₂Cl₂, 0.016 mmol, 1.1 equiv.)and the resulting mixture is stirred at −78° C. for 10 min according tothe procedure described for the synthesis of fluoride 50, to yield,after preparative thin layer chromatography (250 mm silica gel plates,50% EtOAc in hexanes), fluoride 53 (3.0 mg, 50%). R_(f)=0.50 (silicagel, 50% EtOAc in hexanes); [α]²² _(D) −12.4 (c 0.2, CHCl₃); IR (thinfilm) ν_(max) 3408, 2926, 2851, 1732, 1682, 1462, 1384, 1292, 1250,1150, 1068, 974 cm⁻; ¹H NMR (600 MHz, CDCl₃) δ7.04 (s, 1 H, ArH), 6.86(dd, J=17.4, 10.8 Hz, 1 H, CH═CH₂), 6.59 (s, 1 H, CH═CCH₃), 6.05 (d,J=17.5 Hz, 1 H, CH═CH₂), 5.55 (d, J=11.0 Hz, 1 H, CH═CH₂) 5.57-5.51 (m,1 H, C═CHCH₂), 5.25 (d, J=10.0 Hz, 1 H, CHOCO), 4.79 (dd, J 83.8, 10.7Hz, 1 H, CH═CCH₂F), 4.71 (dd, J=83.6, 10.7 Hz, 1 H, CH═CCH₂F), 4.28 (dd,J=10.6, 1.6 Hz, 1 H, CHOH), 3.70 (m, 1 H, CHOH), 3.33-3.25 (m, 1 H,CHOH), 3.16 (qd, J=7.0, 2.1 Hz, 1 H, CH₃CH(C═O)), 2.98 (m, 1 H, OH),2.75-2.66 (m, 1 H, C═CHCH₂CHO), 2.46 (dd, J=14.6, 11.0 Hz, 1 H, CH₂COO),2.37-2.27 (m, 2 H, CH₂C(CH₂OH)═CH), 2.28 (dd, J=14.6, 2.6 Hz, 1 H,CH₂COO), 2.15-2.08 (m, 1 H, C═CHCH₂CHO), 2.11 (s, 3 H, CH═C(CH₃)),1.80-1.64 (m, 3 H), 1.43-1.27 (m, 2 H), 1.34 (s, 3 H, C(CH₃)₂), 1.18 (d,J=6.8 Hz, 3 H, CH(CH₃)), 1.07 (s, 3 H, C(CH₃)₂), 1.03 (d, J=7.0 Hz, 3 H,CH(CH₃)); HRMS (FAB), calcd. for C₂₈H₄₀FNO₅S (M+H⁺), 522.2689 found522.2704.

Epoxide 54 as illustrated in Scheme 9. To a solution of allylic alcohol45 (25.4 mg, 0.049 mmol, 1.0 equiv.) and 4 Å molecular sieves in CH₂Cl₂(0.50 mL) at −40° C. is added dropwise (+)-diethyl-D-tartrate (41 μL,0.59 M in CH₂Cl₂, 0.024 mmol, 0.5 equiv.) followed by titaniumisopropoxide (55 μL, 0.35 M in CH₂Cl₂, 0.019 mmol, 0.4 equiv.). After 1h at that temperature, t-butyl hydroperoxide (22 gL of a 5 M solution indecane, 0.110 mmol, 2.2 equiv.) is added and the reaction mixture isstirred at −30° C. for 2 h. The reaction mixture is then filteredthrough celite into saturated aqueous Na₂SO₄ (10 mL), eluting with EtOAc(10 mL). The resulting biphasic mixture is then stirred for 1 h and thelayers are separated. The aqueous phase is extracted with EtOAc (3×10mL) and the combined organic extracts are dried (MgSO₄) and concentratedunder reduced pressure. Preparative thin layer chromatography (250 mmsilica gel plates, 80% EtOAc in hexanes) furnishes epoxide 54 (13.5 mg,52%). R_(f)=0.23 (silica gel, 80% EtOAc in hexanes); [α]²² _(D) −55.4 (c0.06, CHCl₃); IR (thin film) ν_(max) 3425, 2929, 2862, 1732, 1688, 1456,1367, 1292, 1258, 1195, 1149, 1040, 980 cm⁻¹; ¹H NMR (600 MHz, CHCl₃)δ7.22 (s, 1 H, ArH), 6.62 (s, 1 H, CH═CCH₃), 5.59 (d, J=47.0 Hz, 2 H,ArCH₂F), 5.46 (dd, J=6.7, 3.4 Hz, 1 H, CHOCO), 4.14-4.09 (m, 1 H, CHOH),3.89 (d, J=6.4 Hz, 1 H, OH), 3.76 (bs, 1 H, CHOH), 3.72 (d, J=12.1 Hz, 1H, CH₂OH), 3.56 (dd, J=12.1, 7.5 Hz, 1 H, CH₂OH), 3.33 (qd, J=6.8, 5.3Hz, 1 H, CH₃CH(C═O)), 3.16 (dd, J=6.3, 6.1 Hz, 1 H, C(O)CHCH₂CHO), 2.55(dd, J=14.1, 10.2 Hz, 1 H, CH₂COO), 2.50 (bs, 1 H, OH), 2.41 (dd,J=14.1, 3.1 Hz, 1 H, CH₂COO), 2.11 (s, 3 H, CH═C(CH₃)), 2.10-1.97 (m, 2H, C(O)CHCH₂CHO), 1.91-1.81 (m, 2 H, CH₂C(CH₂OH)), 1.74-1.60 (m, 3 H),1.50-1.30 (m, 2 H), 1.34 (s, 3 H, C(CH₃)₂), 1.18 (d, J=6.8 Hz, 3 H,CH(CH₃)), 1.06 (s, 3 H, C(CH₃)₂), 0.99 (d, J=7.0 Hz, 3 H, C(CH₃)₂); HRMS(FAB), calcd. for C₂₇H₄₀FNO₇S (M+H⁺), 542.2588 found 542.2575.

Epoxide 55 as illustrated in Scheme 9. To a solution of allylic alcohol46 (22 mg, 0.042 mmol, 1.0 equiv.) and 4 Å molecular sieves in CH₂Cl₂(420 μL) at −40° C. is added dropwise (+)-diethyl-D-tartrate (4 μL,0.021 mmol, 0.5 equiv.), followed by titanium isopropoxide (5 μL, 0.016mmol, 0.4 equiv.) and after 1 h at this temperature, t-butylhydroperoxide (18 μL of a 5 M solution in decane, 0.092 mmol, 2.2equiv.) according to the procedure described for the synthesis ofepoxide 54 to yield, after preparative thin layer chromatography (250 mmsilica gel plates, 80% EtOAc in hexanes), epoxide 55 (16 mg, 70%).R_(f)=0.25 (silica gel, 80% EtOAc in hexanes); [α]²² _(D) −44.8 (c 1.4,CHCl₃); IR (thin film) ν_(max) 3435, 2959, 2935, 2877, 1732, 1689, 1534,1459, 1421, 1371, 1338, 1241, 1174, 1039, 980 cm⁻; ¹H NMR (500 MHz,CDCl₃) δ6.51 (s, 1 H, ArH), 6.35 (s, 1 H, CH═CCH₃), 5.40 (dd, J=7.0, 3.0Hz, 1 H, CHOCO), 4.11 (ddd, J=10.0, 6.5, 3.0 Hz, 1 H, CHOH), 4.07 (s, 3H, CH₃O), 3.88 (d, J=6.0 Hz, 1 H, OH), 3.77-3.74 (m, 1 H, CHOH), 3.73(dd, J=12.5, 4.0 Hz, 1 H, CH₂OH), 3.57 (dd, J=12.5, 8.0 Hz, 1 H, CH₂OH),3.32 (qd, J=7.0, 5.0 Hz, 1 H, CH₃CH(C═O)), 3.16 (dd, J=7.0, 5.5 Hz, 1 H,C(O)CHCH₂CHO), 2.54 (dd, J=14.5, 10.0 Hz, 1 H, CH₂COO), 2.50 (bs, 1 H,OH), 2.40 (dd, J=14.5, 3.5 Hz, 1 H, CH₂COO), 2.13 (s, 3 H, CH═C(CH₃)),2.12-2.05 (m, 1 H, C(O)CHCH₂CHO), 2.03-1.95 (m, 2 H), 1.90-1.82 (m, 1 H,CH₂C(CH₂OH)), 1.75-1.60 (m, 2 H), 1.50-1.20 (m, 3 H), 1.35 (s, 3 H,C(CH₃)₂), 1.16 (d, J=7.0 Hz, 3 H, CH(CH₃)), 1.07 (s, 3 H, C(CH₃)₂), 0.99(d, J=7.0 Hz, 3 H); HRMS (FAB), calcd. for C₂₇H₄₁NO₈S (M+Cs⁺), 672.1607found 672.1584.

Fluoride 58 as illustrated in Scheme 9. A solution of triol 54 (5.0 mg,0.009 mmol, 1.0 equiv.) in CH₂Cl₂ (1 mL, 0.01 M) at −78° C. is treatedwith DAST (0.25 mL of a 0.1 M solution in CH₂Cl₂, 0.025 mmol, 1.05equiv.) according to the procedure described for the synthesis offluoride 50, to yield, after preparative thin layer chromatography (250mm silica gel plates, 60% EtOAc in hexanes), fluoride 58 (2.0 mg, 41%).R_(f)=0.22 (silica gel, 50% EtOAc in hexanes); IR (thin film) ν_(max)3402, 2954, 2923, 2853, 1732, 1688, 1462, 1378, 1262, 1185, 1149, 1082,1031, 980 cm⁻¹; ¹H NMR (500 MHz, CHCl₃) δ7.23 (s, 1 H, ArH), 6.63 (s, 1H, CH═CCH₃), 5.60 (d, J=47 Hz, 2 H, ArCH₂F), 5.47 (dd, J=7.0, 3.0 Hz, 1H, CHOCO), 4.39 (dd, J=97.0, 10.5 Hz, 1 H, C(O)CH₂F), 4.30 (dd, J=97.0,10.5 Hz, 1 H, C(O)CH₂F), 4.13 (ddd, J=9.5, 6.5, 3.0 Hz, 1 H, CHOH), 3.75(dd, J=5.0, 5.0 Hz, 1 H, CHOH), 3.74 (d, J=7.0 Hz, 1 H, OH), 3.31 (qd,J=7.0, 6.0 Hz, 1 H, CH₃CH(C═O)), 3.02 (dd, J=6.0, 6.0 Hz, 1 H,CH(O)CH₂CHO), 2.56 (dd, J=14.0, 10.0 Hz, 1 H, CH₂COO), 2.46 (brs, 1 H,OH), 2.42 (dd, J=14.0, 4.0 Hz, 1 H, CH₂COO), 2.13 (s, 3 H, CH═C(CH₃)),2.10-1.97 (m, 3 H), 1.95-1.87 (m, 1 H), 1.90-1.82 (m, 1 H), 1.75-1.63(m, 2 H), 1.50-1.20 (m, 2 H), 1.36 (s, 3 H, C(CH₃)₂), 1.16 (d, J=7.0 Hz,3 H, CH(CH₃)), 1.08 (s, 3 H, C(CH₃)₂), 1.01 (d, J=7.0 Hz, 3 H, C(CH₃)₂);MS (electrospray), calcd. for C₂₇H₃₉F₂NO₆S (M+H⁺) 544, found 544.

Fluoride 59 as illustrated in Scheme 9. A solution of triol 55 (15 mg,0.028 mmol, 1.0 equiv.) in CH₂Cl₂ (280 μL, 0.1 M) at −78° C. is treatedwith DAST (5 μL, 0.038 mmol, 1.4 equiv.) according to the proceduredescribed for the synthesis of fluoride 50, to yield, after preparativethin layer chromatography (250 mm silica gel plates, 50% EtOAc inhexanes), fluoride 59 (4.0 mg, 26%). R_(f)=0.42 (silica gel, 80% EtOAcin hexanes); [α]²² _(D) −29.4 (c 0.33, CHCl₃); IR (thin film) ν_(max)3492, 2960, 2928, 2874, 2865, 1738, 1732, 1693, 1682, 1537, 1462, 1455,1422, 1384, 1241, 1144, 980 cm⁻¹; ¹H NMR (500 MHz, CHCl₃) δ6.52 (s, 1 H,ArH), 6.35 (s, 1 H, CH═CCH₃), 5.41 (dd, J=7.0, 3.5 Hz, 1 H, CHOCO), 4.40(dd, J=111.5, 10.5 Hz, 1 H, CH₂F), 4.30 (dd, J=111.5, 10.5 Hz, 1 H,CH₂F), 4.14 (ddd, J=10.0, 7.0, 3.5 Hz, 1 H, CHOH), 4.08 (s, 3 H, CH₃O),3.80 (d, J=7.0 Hz, 1 H, OH), 3.78 (dd, J=3.5, 3.5 Hz, 1 H, CHOH), 3.31(qd, J=7.0, 5.0 Hz, 1 H, CH₃CH(C═O)), 3.01 (dd, J=7.0, 5.5 Hz, 1 H,C(O)CHCH₂CHO), 2.55 (dd, J=14.5, 10.0 Hz, 1 H, CH₂COO), 2.53 (bs, 1 H,OH), 2.40 (dd, J=14.5, 3.5 Hz, 1 H, CH₂COO), 2.14 (s, 3 H, CH═C(CH₃)),2.12-2.15-1.90 (m, 3 H), 1.73-1.70 (m, 1 H), 1.55-1.24 (m, 5 H), 1.36(s, 3 H, C(CH₃)₂), 1.17 (d, J=6.5 Hz, 3 H, CH(CH₃)), 1.09 (s, 3 H,C(CH₃)₂), 1.00 (d, J=7.0 Hz, 3 H, C(CH₃)₂); HRMS (FAB), calcd. forC₂₇H₄₀FNO₇S (M+Cs⁺), 674.1564 found 674.1594.

Epoxide 57 as shown in Scheme 10. To a solution of allylic alcohol 24(81 mg, 0.151 mmol, 1.0 equiv.) and 4 Å molecular sieves in CH₂Cl₂ (1.25mL) at −40° C. is added dropwise (+)-diethyl-D-tartrate (13 μL, 0.076mmol, 0.5 equiv.), followed by titanium isopropoxide (18 μL, 0.060 mmol,0.4 equiv.) and after 1 h at this temperature, t-butyl hydroperoxide (66μL of a 5 M solution in decane, 0.330 mmol, 2.2 equiv.) and the reactionconducted according to the procedure described for the synthesis ofepoxide 54 to yield, after flash column chromatography (silica gel, 80%EtOAc in hexanes), epoxide 57 (74 mg, 89%). R_(f)=0.34 (silica gel, 80%EtOAc in hexanes); [α]²² _(D) −32.5 (c 0.3, CHCl₃); IR (thin film)ν_(max) 3455, 2959, 2931, 2877, 1733, 1689, 1465, 1377, 1289, 1257,1147, 1040, 979, 912 cm⁻¹; ¹H NMR (600 MHz, CHCl₃) δ6.46 (s, 1 H,CH═CCH₃), 5.48 (dd, J=4.9, 4.7 Hz, 1 H, CHOCO), 4.00 (bm, 1 H, CHOH),3.75 (dd, J=5.6, 3.4 Hz, 1 H, CHOH), 3.71 (d, J=12.5 Hz, 1 H, CH₂OH),3.64 (bs, 1 H, OH), 3.56 (d, J=12.5 Hz, 1 H, CH₂OH), 3.32 (qd, J=6.7,6.7 Hz, 1 H, CH₃CH(C═O)), 3.09 (dd, J=6.3, 6.2 Hz, 1 H, C(O)CHCH₂CHO),2.52 (dd, J=14.3, 9.8 Hz, 1 H, CH₂COO), 2.43 (dd, J=14.3, 3.4 Hz, 1 H,CH₂COO), 2.28 (bs, 1 H, OH), 1.95 (m, 2 H, C(O)CHCH₂CHO), 1.86 (s, 3 H,CH═C(CH₃)), 1.79 (m, 1 H, CH₂C(CH₂OH)), 1.67 (m, 1 H), 1.61 (m, 1 H),1.46 (m, 2 H), 1.33 (s, 3 H, C(CH₃)₂), 1.24 (m, 2 H), 1.15 (d, J=6.8 Hz,3 H, CH(CH₃)), 1.06 (s, 3 H, C(CH₃)₂), 0.98 (d, J=7.0 Hz, 3 H, C(CH₃)₂);HRMS (FAB), calcd for C₂₃H₃₇IO₇ (M+Na⁺), 575.1483 found 575.1462.

Epoxide 56 as illustrated in Scheme 9. A solution of vinyl iodide 57 (20mg, 0.036 mmol, 1.0 equiv.), stannane 8r (29 mg, 0.072 mmol, 1.5 equiv.)and PdCl₂(MeCN)₂ (2.0 mg, 0.004 mmol, 0.1 equiv.) in degassed DMF (360μL, 0.1 M) is stirred at 25° C. for 20 h, according to the proceduredescribed for the synthesis of lactone 18d, to yield, after preparativethin layer chromatography (250 mm silica gel plates, EtOAc), startingvinyl iodide 57 (6 mg, 30%) and macrolactone 56 (10 mg, 51%). R_(f)=0.23(silica gel, 80% EtOAc in hexanes); [α]²² _(D) −60.0 (c 0.14, CHCl₃); IR(thin film) ν_(max) 3414, 2969, 2933, 2872, 1736, 1687, 1458, 1373,1293, 1258, 1150, 980, 914 cm⁻¹; ¹H NMR (500 MHz, CHCl₃) δ6.99 (s, 1 H,ArH), 6.61 (s, 1 H, CH═CCH₃), 5.43 (dd, J=8.0, 3.0 Hz, 1 H, CHOCO), 4.20(ddd, J=9.5, 6.5, 3.0 Hz, 1 H, CHOH), 4.04 (d, J=6.5 Hz, 1 H, OH), 3.77(dd, J=4.0, 4.0 Hz, 1 H, CHOH), 3.74 (dd, J=12.5, 4.0 Hz, 1 H, CH₂OH),3.57 (dd, J=12.5, 8.0 Hz, 1 H, CH₂OH), 3.32 (qd, J=7.0, 4.5 Hz, 1 H,CH₃CH(C═O)), 3.16 (dd, J=7.5, 5.0 Hz, 1 H, C(O)CHH₂CHO), 3.01 (q, J=7.5Hz, 2 H, CH₂CH₃), 2.56 (brs, 1 H, OH), 2.54 (dd, J=14.0, 10.0 Hz, 1 HCH₂COO), 2.38 (dd, J=14.0, 3.0 Hz, 1 H, CH₂COO), 2.14 (ddd, J=15.0, 4.5,3.0 Hz, 1 H, C(O)CHCH₂CHO) 2.11 (s, 3 H, CH═C(CH₃)), 2.02-1.96 (m, 1 H,C(O)CHCH₂CHO), 1.93-1.84 (m, 1 H), 1.74-1.71 (m, 1 H), 1.55-1.25 (m, 5H), 1.40 (t, J=8.0 Hz, 3 H, CH₃CH₂), 1.37 (s, 3 H, C(CH₃)₂), 1.17 (d,J=7.0 Hz, 3 H, CH(CH₃)), 1.08 (s, 3 H, C(CH₃)₂), 1.01 (d, J=7.0 Hz, 3 HC(CH₃)₂); HRMS (FAB), calcd. for C28₆H₄₃NO₇S (M+Na⁺), 560.2658 found560.2640

bis-Silylether 61 as illustrated in Scheme 10. To a solution of triol 57(83 mg, 0.150 mmol, 1.0 equiv.) in DMF (1.5 mL, 0.1 M) is added Et₃N(315 μL, 2.26 mmol, 15 equiv.) followed by TMSCI (152 μL, 1.20 mmol, 8equiv.) and the mixture is stirred at 25° C. for 12 h. The mixture isthen concentrated under reduced pressure and the resulting oil ispartitioned between ether (10 mL) and water (10 mL) and the layers areseparated. The aqueous layer is extracted with ether (3×10 mL) and thecombined extracts are dried (MgSO₄), concentrated under reduced pressureand then filtered through a short plug of silica gel. The resultingfiltrate is concentrated, dissolved in CH₂Cl₂ (5 ml), and silica gel (1g) is added. The resulting slurry is stirred at 25° C. for 12 h,filtered, concentrated and finally passed through a short plug of silicagel to afford the bis-silylether 61 as a foam (103 mg, 98%). R_(f)=0.48(silica gel, 60% Et₂O in hexanes); [α]²² _(D) −19.1 (c 0.23, CHCl₃); IR(thin film) ν_(max) 3408, 2956, 1746, 1698, 1454, 1383, 1250, 1156,1113, 1060, 1021, 985, 898, 841, 752 cm⁻¹; ¹H NMR (500 MHz, CHCl₃) δ6.44(s, 1 H, ArH), 5.37 (dd, J=9.0 Hz, 1 H, CHOCO), 4.01 (dd, J=10.5, 2.5Hz, 1 H, CHOH), 3.86 (d, J=10.0 Hz, 1 H, CHOSi), 3.79 (dd, J=12.5, 4.5Hz, 1 H, CH₂OH), 3.49 (ddd, J=12.5, 10.5, 8.5 Hz, 1 H, CH₂OH), 3.39 (m,1 H, OH), 3.09 (dd, J=10.5, 3.5 Hz, 1 H, CH(O)CH₂CO), 2.97 (qd, J=6.5,4.0 Hz, 1 H, CH₃CH(C═O)), 2.74 (dd, J=16.5, 10.5 Hz, 1 H, CH₂COO), 2.67(dd, J=16.0, 2.5 Hz, 1 H, CH₂COO), 2.18-2.15 (m, 1 H, CH(O)CH₂CHO),1.95-1.82 (m, 2 H), 1.82 (s, 3 H, CH₃C═C), 1.68-1.40 (m, 4 H), 1.24 (m,2 H), 1.18 (s, 3 H, C(CH₃)₂), 1.11 (s, 3 H, C(CH₃)₂), 1.06 (d, J=6.5 Hz,3 H, CH(CH₃)), 0.95 (d, J=7.0 Hz, 3 H, CH(CH₃)), 0.14 (s, 9 H,(CH₃)₃Si), 0.06 (s, 9 H, (CH₃)₃Si); HRMS (FAB), calcd. for C₂₉H₅₃IO₇Si₂(M+Cs⁺), 829.1429 found 829.1459.

Aldehyde 62 as illustrated in Scheme 10. To a suspension of alcohol 61(20 mg, 0.029 mmol, 1.0 equiv.) and 4 Å molecular sieves in CH₂Cl₂ (0.25mL) is added NMO (10 mg, 0.085 mmol, 3.0 equiv.) followed by TPAP (1 mg,0.003 mmol, 0.1 equiv.). The resulting slurry is stirred at 25° C. for40 min and then filtered through a short plug of silica to affordaldehyde 62 (18 mg, 90%). R_(f)=0.66 (silica gel, 60% Et₂O in hexanes);IR (thin film) ν_(max) 2956, 2913, 2851, 1732, 1698, 1454, 1383, 1250,1156, 1113, 1021, 987, 895, 841, 750 cm⁻¹; ⁻¹H NMR (600 MHz, CDCl₃)δ8.84 (s, 1 H, CH═O), 6.51 (s, 1 H, ArH), 5.46 (dd, J=7.9, 3.4 Hz, 1 H,CHOCO), 3.81 (d, J=8.3 Hz, 1 H, CHOSi), 3.32 (dd, J=8.5, 4.2 Hz, 1 H,CHOSi), 3.04 (qd, J=7.1, 7.1 Hz, 1 H CH₃CH(C═O)), 2.65 (dd, J=15.6, 8.3Hz, 1 H, CH₂COO), 2.59 (dd, J=15.6, 4.1 Hz, 1 H, CH₂COO), 2.21 (ddd,J=15.2, 3.8, 3.8 Hz, 1 H, CH(O)CH₂CHO), 2.06-1.97 (m, 2 H), 1.87 (s, 3H, CH₃C═CH), 1.87-1.80 (m, 1 H), 1.62-1.56 (m, 1 H), 1.51-1.41 (m, 2 H),1.27-1.21 (obscured m, 2 H), 1.15 (s, 3H, C(CH₃)₂), 1.08 (s, 3H,C(CH₃)₂), 1.08 (d, J=6.2 Hz, 3 H, CH(CH₃)), 0.96 (d, J=6.9 Hz, 3 H,CH(CH₃)), 0.2 (s, 9 H, (CH₃)₃Si), 0.05 (s, 9 H, (CH₃)₃Si); HRMS (FAB),calcd. for C₂₉H₅₁IO₇Si₂ (M+CS⁺), 827.1272 found 827.1304.

Olefin 63 as illustrated in Scheme 10. Methyltriphenylphosphoniumbromide (104 mg of a mixture with sodium amide (Aldrich), 0.250 mmol,9.7 equiv.) in THF (2.0 mL) is added portionwise to a solution ofaldehyde 62 (18.0 mg, 0.026 mmol, 1.0 equiv.) in THF (0.5 mL) at −5° C.until the completion of the reaction is established by TLC. Saturatedaqueous NH₄Cl (1 mL) is added and the product is extracted with ether(3×2 mL) dried (MgSO₄) and then concentrated under reduced pressure.Flash column chromatography (silica gel, 15% ether in hexanes) furnishesolefin 63 (11.7 mg, 65%). R_(f)=0.50 (silica gel, 20% Et₂O in hexanes);[α]²² _(D) −17.9 (c 0.2, CHCl₃); IR (thin film) ν_(max) 2954, 2923,1747, 1698, 1456, 1382, 1250, 1156, 1113, 1021, 986, 889, 841, 750 cm⁻¹;¹H NMR (500 MHz, CHCl₃) δ6.44 (s, 1 H, ArH), 6.00 (dd, J=17.0, 10.0 Hz,1 H, CH═CH₂), 5.36 (dd, J=9.0, 2.0 Hz, 1 H, CHOCO), 5.29 (dd, J=17.5,1.5 Hz, 1 H, CH₂═CH), 5.14 (dd, J=10.5, 1.5 Hz, 1 H, CH₂═CH), 4.12J=9.0, 5.0 Hz, 1 H, CHOSi), 3.85 (d, J=9.5 Hz, 1 H, CHOSi), 3.04 (qd,J=9.0, 7.0 Hz, 1 H, CH₃CH(C═O)), 2.85 (dd, J=9.5, 4.0 Hz, 1 H,CH(O)CCH═CH₂), 2.73 (dd, J=16.0, 10.0 Hz, 1 H, CH₂COO), 2.65 (dd,J=16.0, 2.5 Hz, 1 H, CH₂COO), 2.12 (ddd, J=15.0, 4.0, 2.0 Hz, 1 H,CH₂CH(O), 1.93-1.78 (3 H, m), 1.84 (s, 3 H, CH═CCH₃), 1.65-1.20 (m, 5H), 1.19 (s, 3 H, C(CH₃)₂), 1.11 (s, 3 H, C(CH₃)₂), 1.08 (d, J=6.5 Hz, 3H, CH(CH₃)), 0.95 (d, J=7.0 Hz, 3 H, CH(CH₃)), 0.14 (s, 9 H, (CH₃)₃Si),0.07 (9 H, s, (CH₃)₃Si), HRMS (FAB), calcd. for C₃₀H₅₃IO₆Si₂ (M+Cs⁺),825.1480 found 825.1450.

Macrolactone 65 as illustrated in Scheme 10. A solution of olefin 63 (15mg, 0.022 mmol, 1.0 equiv.) in EtOH (1.0 mL) is treated with hydrazine(17 μL, 0.500 mmol, 25.0 equiv.) and H₂O₂ (25 μL, 30% w/w in water,0.370 mmol, 16.0 equiv.) and the resulting mixture stirred at 0° C. for3 h. The mixture is then partitioned between ether (4 mL) and water (2mL) and the layers are separated. The aqueous layer is extracted withether (3×4 mL) and the combined organic extracts are dried (MgSO₄) andconcentrated under reduced pressure to give a foam (15.0 mg) which isdissolved in THF (1.5 mL) and treated with HF.pyr. in pyridine/THF (600mL) and the mixture is stirred at 0° C. for 2 h. The reaction mixture isthen quenched with saturated aqueous NaHCO₃ (5 mL) and is extracted withEtOAc (3×3 mL). The combined organic extracts are dried (MgSo₄) andconcentrated under reduced pressure. Flash column chromatography (silicagel, 80% ether in hexanes) furnishes macrolactone 65 (9.4 mg, 75%).R_(f)=0.06 (silica gel, 60% Et₂O in hexanes); [α]²² _(D) −19.3 (c 0.33,CHCl₃); IR (thin film) ν_(max) 3416, 2954, 2926, 2872, 1734, 1689, 1456,1384, 1287, 1256, 1149, 1084, 978, 892 cm⁻¹; ¹H NMR (500 MHz, CHCl₃)δ6.46 (s, 1 H, CH═CCH₃), 5.48 (dd, J=5.0, 5.0 Hz, 1 H, CHOCO), 4.03(brm, 1 H, CHOH), 3.76 (brm, 2 H, CHOH and OH), 3.34 (qd, J=6.5, 6.5 Hz,1 H, CH₃CH(C═O)), 2.73 (dd, J=6.5, 6.5 Hz, 1 H, CH(O)CCH₂CH₃), 2.54 (dd,J=14.5, 10.0 Hz, 1 H, CH₂COO), 2.44 (dd, J=14.5, 8.5 Hz, 1 H, CH₂COO),2.29 (brs, 1 H, OH), 1.96-1.85 (m, 2H), 1.89 (s, 3 H, CH₃C═CH),1.70-1.40 (m, 5 H), 1.31-1.24 (m, 4 H), 1.35 (s, 3 H, C(CH₃)₂), 1.19 (d,J=6.5 Hz, 3 H, CH(CH₃)), 1.07 (s, 3 H, C(CH₃)₂), 0.99 (d, J=7.0 Hz, 3 H,CH(CH₃)), 0.91 (t, J=7.5 Hz, 3 H, CH₃CH₂); HRMS (FAB), calcd. forC₂₄H₃₉IO₆ (M+Cs⁺), 683.0846 found 683.0870.

Macrolactone 66 as illustrated in Scheme 10. A solution of vinyl iodide65 (9.4 mg, 0.017 mmol, 1.0 equiv.), stannane 8j (10 mg, 0.036 mmol, 2.1equiv.) and PdCl₂(MeCN)₂ (1.0 mg, 0.004 mmol, 0.2 equiv.) in degassedDMF (250 μL, 0.07 M) is stirred at 25° C. for 15 h, according to theprocedure described for the synthesis of macrolactone 18d, to yield,after preparative thin layer chromatography (250 mm silica gel plates,EtOAc) macrolactone 66 (4.6 mg, 52%). R_(f)=0.40 (silica gel, 80% EtOAcin hexanes); [α]²² _(D) −30.0 (c 0.17, CHCl₃); IR (thin film) ν_(max)3432, 2967, 2933, 2872, 1736, 1689, 1458, 1384, 1256, 1151, 1067, 1038,979, 905, 733 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ7.23 (s, 1 H, ArH), 6.62(s, 1 H, CH═CCH₃), 5.59 (d, J=47.1 Hz, 2 H, CH₂F), 5.46 (dd, J=6.3, 3.7Hz, 1 H, CHOCO), 4.15 (d, J=8.8 Hz, 1 H, CHOH), 3.98 (brs, 1 H, OH),3.77 (brs, 1 H, CHOH), 3.35 (qd, J=6.6, 4.8 Hz, 1 H, CH₃CH(C═O)), 2.82(dd, J=6.1, 6.1 Hz, 1 H, CH(O)CCH₂CH₃), 2.56 (dd, J=14.0, 9.9 Hz, 1 H,CH₂COO), 2.48 (brs, 1 H, OH), 2.41 (dd, J=14.0, 3.0 Hz, 1 H, CH₂COO),2.13 (s, 3 H, CH═C(CH₃)), 2.04 (ddd, J=15.1, 5.9,4.0 Hz, 1 H,CH₂CH(O)CHCH₂), 2.00-1.94 (m, 1 H, CH₂CH(O)CHCH₂), 1.78-1.24 (m, 7 H),1.36 (s, 3 H, C(CH₃)₂), 1.17 (d, J=7.0 Hz, 3 H, CH(CH₃)), 1.07 (s, 3 H,C(CH₃)₂), 1.00 (d, J=7.0 Hz, 3 H, CH(CH₃)); HRMS )FAB calcd. forC₂₈H₄₂FNO₆S (M+Cs⁺), 672.1771 found 672.1793.

Macrolactone 67 as illustrated in Scheme 10. A solution of vinyl iodide65 (11 mg, 0.020 mmol, 1.0 equiv.), stannane 8p (14 mg, 0.034 mmol, 1.7equiv.) and PdCl₂(MeCN)₂ (1.0 mg, 0.004 mmol, 0.2 equiv.) in degassedDMF (250 μL, 0.08 M) is stirred at 25° C. for 20 h, according to theprocedure described for the synthesis of macrolactone 18d, to yield,after preparative thin layer chromatography (250 mm silica gel plates,EtOAc) macrolactone 67 (8.5 mg, 79%). R_(f)=0.68 (silica gel, Et₂O);[α]²² _(D) −44.7 (c 0.08 CHCl₃); IR (thin film) ν_(max) 3442, 2964,2934, 1732, 1683, 1536, 1461, 1422, 1384, 1241, 1150, 1070, 979, 906,732 cm⁻; ¹H NMR (500 MHz, CDCl₃) δ6.52 (s, 1 H, ArH), 6.36 (s, 1 H,CH═CCH₃), 5.41 (dd, J=7.0, 3.3 Hz, 1 H, CHOCO), 4.15 (ddd, J=10.3, 7.0,3.7 Hz, 1 H, CHOH), 4.08 (s, 3 H, OCH₃), 3.99 (brd, J=6.3 Hz, 1 H, OH),3.77 (brm, 1 H, CHOH), 3.34 (qd, J=6.6, 4.8 Hz, 1 H, CH₃CH(C═O)), 2.81(dd, J=6.6, 5.9 Hz, 1 H, CH(O)CCH₂CH₃), 2.55 (dd, J=14.2, 10.1 Hz, 1 H,CH₂COO), 2.52 (brs, 1 H, OH), 2.39 (dd, J=14.0, 2.9 Hz, 1 H, CH₂COO),2.14 (s, 3 H, CH═C(CH₃)), 2.05 (ddd, J=15.1, 5.5, 4.0 Hz, 1 H,CH₂CH(O)CHCH₂), 1.98-1.92 (m, 1 H, CH₂CH(O)CHCH₂), 1.80-1.70 (m, 2 H),1.58-1.39 (m, 5 H), 1.30-1.24 (m, 2 H), 1.17 (d, J=7.0 Hz, 3 H,CH(CH₃)), 1.08 (s, 3 H, C(CH₃)₂), 1.00 (d, J=7.0 Hz, 3 H, CH(CH₃)), 0.91(t, J=7.4 Hz, 3 H, CH₃CH₂); HRMS (FAB), calcd. for C₂₈H₄₃NO₇S (M+Cs⁺),670.1815 found 670.1837.

Macrolactone 68 as illustrated in Scheme 10. A solution of vinyl iodide65 (5.8 mg, 0.011 mmol, 1.0 equiv.), stannane 8r (10 mg, 0.025 mmol, 2.3equiv.) and PdCl₂(MeCN)₂ (1.0 mg, 0.004 mmol, 0.3 equiv.) in degassedDMF (100 μL, 0.1 M) is stirred at 25° C. for 23 h, according to theprocedure described for the synthesis of macrolactone 18d, to yield,after preparative thin layer chromatography (250 mm silica gel plates,EtOAc) macrolactone 68 (3.7 mg, 65%). R_(f)=0.45 (silica gel, Et₂O);[α]²² _(D) −33.3 (c 0.09, CHCl₃); IR (thin film) ν_(max) 3406, 2954,2924, 2872, 1736, 1692, 1454, 1384, 1254, 1150, 1071, 979 cm⁻¹; ¹H NMR(500 MHz, CDCl₃) δ6.99 (s, 1 H, ArH), 6.60 (s, 1 H, CH═CCH₃), 5.42 (dd,J=7.9, 3.1 Hz, 1 H, CHOCO), 4.33 (brs, 1 H, CHOH), 4.24 (brd, J=9.6 Hz,1 H, OH), 3.76 (brm, 1 H, CHOH), 3.32 (qd, J=6.8, 4.3 Hz, 1 H,CH₃CH(C═O)), 3.01 (q, J=7.6 Hz, 2 H, ArCH₂CH₃), 2.82 (dd, J=7.4, 4.8 Hz,1 H, CH(O)CH₂), 2.60 (brs, 1 H, OH), 2.54 (dd, J=13.6, 10.3 Hz, 1 H,CH₂COO), 2.35 (dd, J=14.0, 2.9 Hz, 1 H, CH₂COO), 2.10-2.05 (obscured m,1 H, CH₂CH(O)), 2.09 (s, 3 H, CH═C(CH₃)), 1.96-1.90 (m, 1 H,CH₂CH(O)CHCH₂), 1.80-1.67 (m, 2 H), 1.66-1.25 (m, 7 H), 1.38 (s, 3 H,C(CH₃)₂), 1.16 (d, J=7.0 Hz, 3 H CH(CH₃)), 1.07 (s, 3 H, C(CH₃)₂), 1.00(d, J=7.0 Hz, 3 H CH(CH₃)), 0.92 (t, J=7.4 Hz, 3 H, CH₃CH₂), 0.91 (t,J=7.5 Hz, 3 H, CH₃CH₂); HRMS (FAB), calcd. for C₂₉H₄₅NO₆S (M+Cs⁺),668.2022 found 668.2042.

Tubulin Polymerization and Cytotoxicity Assays

Tubulin polymerization is determined by the filtration-colorimetricmethod, developed by Bollag et Cancer Res. 1995, 55, 2325-2333. Purifiedtubulin (1 mglmL) is incubated at 37° C. for 30 minutes in the presenceof each compound (20 mM) in MEM buffer [(100 mM2-(N-morpholino)ethanesulfonic acid, pH 6.75, 1 mM ethylene glycolbis(β-aminoethyl ether), N,N,N′N′-tetraacetic acid, and 1 mM MgCl₂]; themixture is then filtered to remove unpolymerized tubulin by using a96-well Millipore Multiscreen Durapore hydrophilic 0.22 μm pore sizefiltration plate; the collected polymerized tubulin is stained withamido black solution and quantified by measuring absorbance of the dyedsolution on a Molecular Devices Microplate Reader. The growth of allcell lines is evaluated by quantitation of the protein in 96-well platesas described previously. Briefly, 500 cells are seeded in each well ofthe plates and incubated with the various concentrations of theepothilones at 37° C. in a humidified 5% CO₂ atmosphere for four days.After cell fixation with 50% trichloroacetic acid, the optical densitycorresponding to the quantity of proteins is measured in 25 mM NaOHsolution (50% methanol:50% water) at a wavelength of 564 nm. The IC50 isdefined as the dose of drug required to inhibit cell growth by 50%.

Scheme 11 is shown using conditions described in Nicolaou et al. J. Am.Chem. Soc., 1997, 119, 7974-7991 and those as indicated in thedescription of Scheme 11 above.

Vinyl iodide 7002 as illustrated in Scheme 11. Diiodide 7001 (1 equiv.;from 57) and sodium cyanoborohydride (10 equiv.) are dissolved inanhydrous HMPA (0.2 M) and the resulting mixture heated at 45-50° C. for48 h. After cooling to room temperature, water is added and the aqueousphase extracted four times with ethyl acetate. The combined organicfractions are dried (Na₂SO₄) and passed through a short plug of silicagel to remove traces of HMPA (eluting with 50% ethyl acetate inhexanes). Following evaporation of solvents, the residue is purified bypreparative thin layer chromatography (eluting with 50% ethyl acetate inhexanes) to provide pure vinyl iodide 7002 (84%).

What is claimed is:
 1. A compound represented by the followingstructure:

wherein the waved bond indicates that bond “a” is present either in thecis or in the trans form; (i) R₃ is a radical selected from the groupconsisting of hydrogen; lower alkyl (C₁-C₆); —CH═CH₂; —C≡CH; —CH₂F;—CH₂Cl; —CH₂—OH; —CH₂—O—(C₁-C₆ alkyl); and —CH₂—S—(C₁-C₆ alkyl); R₄ andR₅ are independently selected from the group consisting of hydrogen,methyl, and a protecting group; and R₁ is a radical selected from thefollowing structures:

(ii) and, if R₃ is lower alkyl (C₁-C₆); —CH═CH₂; —C≡CH; —CH₂F; —CH₂Cl;—CH₂—OH; —CH₂—O—(C₁-C₆ alkyl); and —CH₂—S—(C₁-C₆ alkyl); and the othersymbols except R₁ have the same meanings given above, R₁ can also be aradical selected from the following structures:

(iii) and, if R₃ is lower alkyl (C₂-C₆); —CH═CH₂; —C≡CH; —CH₂F; —CH₂Cl;—CH₂—OH; —CH₂—O—(C₁-C₆ alkyl); and —CH₂—S—(C₁-C₆ alkyl); and the othersymbols except R₁ have the same meanings given above under (i), R₁ canalso be a moiety of the formula

or a salt thereof where one or more salt-forming groups are present. 2.A compound of the formula I according to claim 1 wherein: (i) R₃ is aradical selected from the group consisting of hydrogen; lower alkyl(C₁-C₆); —CH═CH₂; —C≡CH; —CH₂F; —CH₂Cl; —CH₂—OH; —CH₂—O—(C₁-C₆ alkyl);and —CH₂—S—(C₁-C₆ alkyl); R₄ and R₅ are independently selected from thegroup consisting of hydrogen, methyl, and a protecting group; and R₁ isa radical selected from the following structures:

or a salt thereof where one or more salt-forming groups are present. 3.A compound of the formula I according to claim 1 wherein: (i) R₃ is aradical selected from the group consisting of hydrogen; lower alkyl(C₁-C₆); —CH═CH₂; —C≡CH; —CH₂F; —CH₂Cl; —CH₂—OH; —CH₂—O—(C₁-C₆ alkyl);and —CH₂—S—(C₁-C₆ alkyl); R₄ and R₅ are independently selected from thegroup consisting of hydrogen, methyl, and a protecting group; and R₁ isa radical selected from the following structure:

or a salt thereof where one or more salt-forming groups are present. 4.A compound of the formula I according to claim 1 wherein: (i) R₃ is alower alkyl (C₁-C₆); —CH═CH₂; —C≡CH; —CH₂F; —CH₂Cl; —CH₂—OH;—CH₂—O—(C₁-C₆ alkyl); and —CH₂—S—(C₁-C₆ alkyl); R₄ and R₅ areindependently selected from the group consisting of hydrogen, methyl,and a protecting group; and R₁ is a radical selected from the followingstructure:

or a salt thereof where one or more salt-forming groups are present. 5.A compound of the formula I according to claim 1 wherein: (i) R₃ is alower alkyl (C₂-C₆); —CH═CH₂; —C≡CH; —CH₂F; —CH₂Cl; —CH₂—OH;—CH₂—O—(C₁-C₆ alkyl); and —CH₂—S—(C₁-C₆ alkyl); R₄ and R₅ areindependently selected from the group consisting of hydrogen, methyl,and a protecting group; and R₁ is a radical of the formula


6. A compound of the formula Id

wherein A is fluoromethyl, methoxy, methylthio or ethenyl (—CH═CH₂) andD is hydrogen, fluoro, hydroxy or methyl.
 7. A compound according toclaim 1, selected from the group consisting of a compound of thefollowing formulae:

or a salt thereof where one or more salt-forming groups are present. 8.A pharmaceutical composition comprising a compound according to any oneof claims 1-4, 5, and 6 and a pharmaceutically acceptable carrier.
 9. Amethod of treatment of a warm-blooded animal suffering a proliferativedisease and that is in need of such treatment, comprising administeringa compound of the formula I, or a pharmaceutically acceptable saltthereof, according to claim 1 to said warm-blooded animal in an amountthat is sufficient for said treatment, said proliferative disease beingselected from the group consisting of lung cancer, prostate cancer,cancer of the intestine, epidermoid tumor, breast cancer, bladdercancer, pancreas cancer, brain cancer, and melanoma.
 10. A method forthe synthesis of a compound of the formula I, in claim 1, comprising a)coupling a iodide of the formula II,

wherein R₂, R₃, R₄, R₅, a, b and c and the waved bond have the meaningsgiven under formula I in claim 1, with a tin compound of the formulaIII, R₁—Sn(R)₃  (III) wherein R₁ has the meanings given under formula Iand R is lower alkyl, especially methyl or n-butyl, or b) coupling a tincompound of the formula IV,

wherein R₂, R₃, R₄, R₅, a, b and c and the waved bond have the meaningsgiven under formula I, with a iodide of the formula V, R₁—I  (V) whereinR₁ has the meanings given under formula I in claim 1; and, if desired, aresulting compound of the formula I is converted into a differentcompound of the formula I, a resulting free compound of the formula I isconverted into a salt of a compound of the formula I, and/or a resultingsalt of a compound of the formula I is converted into a free compound ofthe formula I or into a different salt of a compound of the formula I,and/or a stereoisomeric mixture of compounds of formula I is separatedinto the corresponding isomers.
 11. A method for synthesizing anepothilone E or F of the formula IX,

wherein Q is Hydrogen or methyl, the method comprising the followingstep: epoxidizing a compound of the formula X to the compound of theformula IX in the presence of an epoxide, the compound of the formula Xbeing represented as follows,

wherein Q is hydrogen or methyl.
 12. A compound represented by thefollowing structure:

wherein the waved bond indicates that bond “a” is present either in thecis or in the trans form; and R₂ is absent or oxygen; “a” can be eithera single or double bond; “b” can be either absent or a single bond; and“c” can be either absent or a single bond, with the proviso that if R₂is oxygen then “b” and “c” are both a single bond and “a” is a singlebond; if R₂ is absent then “b” and “c” are absent and “a” is a doublebond; and if “a” is a double bond, then R₂, “b” and “c” are absent; R₃is a radical selected from the group consisting of lower alkyl (C₁-C₆);—CH═CH₂; —C≡CH; —CH₂F; —CH₂Cl; —CH₂—OH; —CH₂—O—(C₁-C₆ alkyl); and—CH₂—S—(C₁-C₆ alkyl); R₄ and R₅ are independently selected from thegroup consisting of hydrogen, methyl, and a protecting group; and R₁ isa radical selected from the following structures:

or a salt thereof where one or more salt-forming groups are present. 13.A compound according to claim 12 represented by the following structure:

wherein A is a radical selected from the group consisting offluoromethyl, and methylthio; and D is a radical selected from the groupconsisting of hydrogen, fluoro, hydroxy, and methyl.
 14. A compoundaccording to claim 13 represented by the following structure:


15. A compound represented by the following structure:

wherein A is radical selected from the group consisting of fluoromethyl,methoxy, methylthio, and ethenyl (—CH═CH₂); and D is a radical selectedfrom the group consisting of hydrogen, fluoro, hydroxy, and methyl; or asalt thereof where one or more salt-forming groups are present.
 16. Acompound according to claim 15 represented by the following structure:


17. A compound represented by the following structure: